"
Template:Team:Uppsala/JS/notebook
From 2013.igem.org
(Difference between revisions)
(Blanked the page) |
|||
| (95 intermediate revisions not shown) | |||
| Line 1: | Line 1: | ||
| + | <html> | ||
| + | <script type="text/javascript"> | ||
| + | var month = 8; | ||
| + | var year = new Date().getFullYear(); | ||
| + | var cal_table = ''; | ||
| + | var eventString; | ||
| + | var days_in_month = new Array(31,28,31,30,31,30,31,31,30,31,30,31); | ||
| + | var days_with_dirys = new Array('t2013610','t2013612','t2013613','t2013617','t2013618','t2013619','t2013620','t2013621','t2013622','t2013623','t2013624','t2013625','t2013626','t2013627','t2013628','t2013629','t2013630','t2013630','t201371','t201372','t201373','t201374','t201375','t201376','t201377','t201379','t201378','t201379','t2013710','t2013711','t2013712','t2013713','t2013714','t2013715','t2013716','t2013717','t2013718','t2013719','t2013720','t2013721','t2013722','t2013723','t2013724','t2013725','t2013726','t2013727','t2013728','t2013729','t2013730','t2013731','t201381','t201382','t201383','t201384','t201385','t201386','t201387','t201388','t201389','t2013810','t2013811','t2013812','t2013813','t2013814','t2013815','t2013816','t2013817','t2013818','t2013819','t2013820','t2013821','t2013822','t2013823','t2013824','t2013825','t2013826','t2013827','t2013828','t2013829','t2013830','t2013831','t201391','t201392','t201393','t201394','t201395','t201396','t201397','t201398','t201399','t2013910','t2013911','t2013912','t2013917','t2013918','t2013919','t2013920','t2013921','t2013924','t2013927','t2013928','t2013101','t2013102','t2013104','t20131011','t20131012','t20131013'); | ||
| + | var inputs = 106; | ||
| + | |||
| + | Date.prototype.getWeek = function() | ||
| + | { | ||
| + | var onejan = new Date(this.getFullYear(),0,1); | ||
| + | return Math.ceil((((this - onejan) / 86400000) + onejan.getDay()+1)/7); | ||
| + | } | ||
| + | function calendar() | ||
| + | { | ||
| + | var m = month+1; | ||
| + | var date = new Date(); | ||
| + | var today = date.getDate(); | ||
| + | cal_table = ''; | ||
| + | var eventString; | ||
| + | |||
| + | if(year<=200) | ||
| + | { | ||
| + | year += 1900; | ||
| + | } | ||
| + | months = new Array('January', 'February', 'March', 'April', 'May', 'June', 'July', 'August', 'September', 'October', 'November', 'December'); | ||
| + | var weekDay = new Array('Mon', 'Tue', 'We', 'Thu', 'Fri', 'Sat', 'Sun'); | ||
| + | days_in_month = new Array(31,28,31,30,31,30,31,31,30,31,30,31); | ||
| + | |||
| + | if(year%4 == 0) | ||
| + | { | ||
| + | if( year!=1900 ) | ||
| + | { | ||
| + | days_in_month[1]=29; | ||
| + | } | ||
| + | } | ||
| + | |||
| + | total = days_in_month[month]; | ||
| + | var date_today = months[month]+' '+year; | ||
| + | |||
| + | cal_table = cal_table + '<table class="cal_calendar" ><tbody id="cal_body"><tr class="head"><th class="arrow"> <input id="left" type="image" onclick="left_arrow()" src="https://static.igem.org/mediawiki/2013/a/a1/Arrowleft.png"></th><th colspan="6" >'+date_today+'</th><th class="arrow"><input id="right" type="image" onclick="right_arrow()" src="https://static.igem.org/mediawiki/2013/0/0b/Arrowright.png"></th></tr>'; | ||
| + | cal_table = cal_table + '<tr class="cal_d_weeks"><th>Mon</th><th>Tue</th><th>Wed</th><th>Thu</th><th>Fri</th><th>Sat</th><th>Sun</th><th>Week</th></tr><tr>'; | ||
| + | week = 0; | ||
| + | |||
| + | var first_week_day = new Date(year, month, 0).getDay(); | ||
| + | |||
| + | for(week_day = 0; week_day < first_week_day; week_day++) | ||
| + | { | ||
| + | cal_table = cal_table +'<td class="cal_days_bef_aft"></td>'; | ||
| + | week++; | ||
| + | } | ||
| + | |||
| + | for(i=1;i<=total;i++) | ||
| + | { | ||
| + | if(week==0) | ||
| + | { | ||
| + | cal_table = cal_table + '<tr>'; | ||
| + | } | ||
| + | |||
| + | if(year == date.getFullYear()) | ||
| + | { | ||
| + | if(month == date.getMonth()){ | ||
| + | if(today==i){ | ||
| + | cal_table = cal_table +'<td id="'+'t'+year+m+i+'" class="cal_today"><input type="submit" onclick="showNotes(year, month, value)" value="'+i+'"></td>'; | ||
| + | } | ||
| + | else{ | ||
| + | cal_table = cal_table +'<td id="'+'t'+year+m+i+'" class="not_today"><input type="submit" onclick="showNotes(year, month, value)" value="'+i+'"></td>'; | ||
| + | } | ||
| + | } | ||
| + | else{ | ||
| + | cal_table = cal_table +'<td id="'+'t'+year+m+i+'" class="not_today"><input type="submit" onclick="showNotes(year, month, value)" value="'+i+'"></td>'; | ||
| + | } | ||
| + | } | ||
| + | |||
| + | else | ||
| + | { | ||
| + | cal_table = cal_table +'<td id="'+'t'+year+m+i+'" class="not_today"><input type="submit" onclick="showNotes(year, month, value)" value="'+i+'"></td>'; | ||
| + | } | ||
| + | |||
| + | week++; | ||
| + | markerNotes(year, m, i); | ||
| + | if(week==7) | ||
| + | { | ||
| + | var weeknr = new Date(year, month, i-1).getWeek(); | ||
| + | cal_table = cal_table + '<td class="week"><input type="submit" onclick="showNotesOfweek(year, value)" value="'+weeknr+'"></td>'; | ||
| + | cal_table = cal_table + '</tr>'; | ||
| + | week=0; | ||
| + | } | ||
| + | |||
| + | } | ||
| + | |||
| + | for(i=1;week!=0;i++) | ||
| + | { | ||
| + | cal_table = cal_table + '<td class="cal_days_bef_aft"></td>'; | ||
| + | week++; | ||
| + | |||
| + | if(week==7) | ||
| + | { | ||
| + | if(weeknr == 53) | ||
| + | { | ||
| + | var weeknr = 1; | ||
| + | } | ||
| + | else | ||
| + | { | ||
| + | var weeknr = weeknr+1; | ||
| + | } | ||
| + | cal_table = cal_table + '<td class="week"><input type="submit" onclick="showNotesOfweek(year, value)" value="'+weeknr+'"></td>'; | ||
| + | cal_table = cal_table + '</tr>'; | ||
| + | week=0; | ||
| + | } | ||
| + | } | ||
| + | cal_table = cal_table + '</tbody></table>'; | ||
| + | } | ||
| + | |||
| + | |||
| + | function left_arrow() | ||
| + | { | ||
| + | month = month-1; | ||
| + | if(month == -1) | ||
| + | { | ||
| + | year = year-1; | ||
| + | month = 11; | ||
| + | } | ||
| + | calendar(); | ||
| + | document.getElementById('calendar').innerHTML = cal_table; | ||
| + | } | ||
| + | |||
| + | function right_arrow() | ||
| + | { | ||
| + | month = month+1; | ||
| + | if(month == 12) | ||
| + | { | ||
| + | year = year+1; | ||
| + | month = 0; | ||
| + | } | ||
| + | calendar(); | ||
| + | document.getElementById('calendar').innerHTML = cal_table; | ||
| + | } | ||
| + | |||
| + | function showNotes(y, m, day) | ||
| + | { | ||
| + | var m = month+1; | ||
| + | |||
| + | eventString = '<div class="eventBox" id="'+'d'+y+m+day+'" onclick="diry('+y+', '+m+', '+day+');"><h1>'+y+'-'+m+'-'+day+'</h1></div>'; | ||
| + | document.getElementById('file_holder').innerHTML = eventString; | ||
| + | document.getElementById('calendar').style.marginBottom="350px"; | ||
| + | } | ||
| + | |||
| + | |||
| + | function showNotesOfweek(y, wnr) | ||
| + | { | ||
| + | var d = new Date(y, 0, 0); | ||
| + | if(y == 2012 ){ | ||
| + | wnr++; | ||
| + | } | ||
| + | var w = d.getTime() + 604800000 * (wnr-1); | ||
| + | var m = new Date(w).getMonth()+1; | ||
| + | var y = new Date(w).getFullYear(); | ||
| + | var day = new Date(w).getDate(); | ||
| + | |||
| + | var e = new Date(w).getDay(); | ||
| + | day = day - e+1; | ||
| + | |||
| + | eventString = '<div class="eventBox" id="'+'d'+y+m+day+'" onclick="diry('+y+', '+m+', '+day+');"><h1>'+y+'-'+m+'-'+day+'</h1></div>'; | ||
| + | |||
| + | for(i=1;i<7;i++) | ||
| + | { | ||
| + | if(days_in_month[m-1] == day ) | ||
| + | { | ||
| + | day = 0; | ||
| + | m++; | ||
| + | if(m>12) | ||
| + | { | ||
| + | y++; | ||
| + | var d = new Date(y, 0, 0); | ||
| + | var w = d.getTime() + 604800000 * (wnr-1); | ||
| + | var m = 1; | ||
| + | } | ||
| + | |||
| + | } | ||
| + | day++; | ||
| + | eventString = eventString + '<div class="eventBox" id="'+'d'+y+m+day+'" onclick="diry('+y+', '+m+', '+day+');"><h1>'+y+'-'+m+'-'+day+'</h1></div>'; | ||
| + | } | ||
| + | document.getElementById('calendar').style.marginBottom="350px"; | ||
| + | document.getElementById('file_holder').innerHTML = eventString; | ||
| + | eventString = ''; | ||
| + | } | ||
| + | function markerNotes(y, m, d) | ||
| + | { | ||
| + | var id='t'+y+m+d; | ||
| + | |||
| + | for(c = 0; c<=inputs; c++) | ||
| + | { | ||
| + | if(days_with_dirys[c] == id) | ||
| + | { | ||
| + | cal_table = cal_table + '<style type="text/css">#'+id+'{border: 2px solid #0089d1;}</style>'; | ||
| + | } | ||
| + | } | ||
| + | } | ||
| + | |||
| + | function diry(y, m, d) | ||
| + | { | ||
| + | var ds = ''; //diryString | ||
| + | var id = 'd'+y+m+d; | ||
| + | //var ds = y+'-'+m+'-'+d; | ||
| + | if(id == 'd2013630') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Sunday 2013-06-30</h1><b>Name of participants:</b> Magnus B, Alexander W, Nils A, Mikael S, Anders E<br><br><h2>Ongoing constructs:</h2><b>E. coli:</b><br>75. pEL3A15 - CrtB<br>25. pSB1C3-CrtY<br>27. pEL3K16-red<br>51.pEL3C18-red<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br>42. pSB4S15-red<br>60. PK401 dam<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69.pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-Cp44-B0032-BFP<br>77. pSB4C15<br><br><b>L. reuteri:</b><br>Lb1. CF48-pLR581<br>Lb2. 1063-pLUL631<br>Lb3. DSM 20016-pVs2<br>Lb4. 100-23-noplasmid<br>Lb6. DSM 20016-pLUL63TsA8<br>Lb7. DSM 20016-pGFP<br>Lb8. 100-23-pGT232<br>Lb12. DSM 20016-noplasmid<br>Lb14. DSM 20016-pLUL631<br><br><b>L. plantarum:</b><br>Lb5. 256-rifR-pAMβ1<br><br><b>L. lactis:</b><br>Lb13. MG1363-pJP059<br>Lb15. unknown-pGus<br>Lb16. MG1363-noplasmid<br><br>Lb10. 256-noplasmid<br>Lb11. 36E-noplasmid<br>Lb9. JH2-2 pAMβ1<br><br><h2>Todays work:</h2><br><b>Transformation</b><br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>77. pSB4C15-red<br>75. pEL3A15 - CrtB<br><br><b>Digest</b><br>27. pEL3K16-red<br>51.pEL3C18-red<br><br><b>Ligation</b><br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>77. pSB4C15-red<br>We made a transformation with one sample of 77 from today, and one from yesterday to try to see if the problem yesterday laid with the transformation or the assembly.<br><br><b>O/N Screening</b><br>25. pSB1C3-CrtY<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br><br><h2>Results</h2><br><b>Re-streak</b><br>Lb13. Lc. lactis MG1363 pJP059: Good growth<br>Lb16. Lc. lactis MG1363 noplasmid: Good growth<br>Lb15. Lc. lactis MG1363-pGUS: Good growth<br><br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF failed<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF failed<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF failed<br>75. pEL3A15 - CrtB failed<br><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013629') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-06-29</h1><br><b>Name of participants:</b> Magnus B, Alexander W, Nils A, Anders E, Mikael S<br><br><h2>Ongoing constructs:</h2><b>L. reuteri:</b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>12. DSM 20016-noplasmid<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-noplasmid<br>11. 36E-noplasmid<br><br><b>E. faecalis:</b><br>9. JH2-2 pAMβ1<br><br><b>L. lactis:</b><br>13. MG1363-pJP059<br>15. unknown-pGus<br>16. MG1363-noplasmid<br><br><b>E.coli (strain D5α):</b><br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br>23. pSB1C3-CrtB<br>24.B0034-CrtI<br>38. pEL3A16-red<br>42. pSB4S15-red<br>57.B0034-Idi<br>75. pEL3A15-CrtB<br>76. pEL3K16-CrtI<br>60. PK401 dam<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69.pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-Cp44-B0032-BFP<br>77. pSB4C15<br><br><h2>Todays work</h2><b>PCR purification:</b><br>CP1<br>CP8<br>CP11<br>CP29<br>CP30<br>CP41<br>Cp44<br><br><b>Digestion:</b><br>27. pEL3K16-red<br>51.pEL3C18-red<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69.pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-Cp44-B0032-BFP<br>77. pSB4C15 (Assembly of 42. pSB4S15 and precut CmR)<br><br><b>Ligation:</b><br>24.3 B0034-CrtI -> 27. pEL3K16-red<br>23.2 pSB1C3-CrtB -> 38. pEL3A16-red<br><br><b>Transformation:</b><br>75. pEL3A15 – CrtB*<br>76. pEL3K16 – CrtI* <br>*Cancelled, wrong solution was used -> redo tomorrow.<br><br><b>Re-streak:</b><br>1.2 pSB1C3-red (from frozen stock)<br>7. pSB3K3 (”)<br>60. PK401 dam<br><br><b>Plasmid preparation:</b><br>42. pSB4S15-red<br><br><b>O/N Screening PCR:</b><br>25. pSB1C3-CrtY<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br>(19,20,21(with DMSO dilution series(7,9,12))<br><br><b>PCR (screening?):</b><br>57.B0034-Idi<br>IspA<br><br><b>Gel electrophoresis:</b><br>57.B0034-Idi<br>IspA<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br>38. pEL3A16-red (digest)<br>23.1 pSB1C3-CrtB (digest)<br>23.2 pSB1C3-CrtB (digest)<br>27. pEL3K16-red (digest)<br>24.3 B0034-CrtI (digest)<br>24.4 B0034-CrtI (digest)<br><br><h2>Results</h2><b>Today:</b><br><b>Plasmid preparation:</b><br>42.1 pSB4S15-red klon 1: 27.4 ng/µL<br>42.2 pSB4S15-red klon 2: 21.8 ng/µL<br>42.3 pSB4S15-red klon 3: 24.3 ng/µL<br>42.4 pSB4S15-red klon 4: 25.3 ng/µL<br><br><b>Gel electrophoresis:</b><br>57.B0034-Idi: still not good after three attempts<br>IspA: eventually good<br>38. pEL3A16-red (digest): failed, redo subcloning tomorrow <br>23.1 pSB1C3-CrtB (digest): failed, redo subcloning tomorrow<br>23.2 pSB1C3-CrtB (digest): failed, redo subcloning tomorrow<br>27. pEL3K16-red (digest): succeded <br>24.3 B0034-CrtI (digest): failed, redo subcloning tomorrow<br>24.4 B0034-CrtI (digest): failed, redo subcloning tomorrow<br><br><b>Transformation:</b><br>75. pEL3A15 - CrtB failed<br><br><b>PCR purification (of promotors):*</b><br>CP1: 40 ng/µl<br>CP8: 41,3 ng/µl<br>CP11: 46,7 ng/µl<br>CP29: 40 ng/µl<br>CP30: 58 ng/µl<br>CP41: 41,9 ng/µl<br>Cp44: 42,3 ng/µl<br><br>*PCR-purification of the promotors may have washed away the promotors themselves, as they are almost as short as the primers that purification is meant to remove, on the other hand, the DNA-concentration of the purified product was measured to around 40 ng/µl. → redo all promotor constructs (not 77) from annealing of R and F (without PCR-cleaning).<br><br><b>Previous day:</b><br><b>ON screening:</b><br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF: failed -> redo today with dilutions<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF: failed -> redo today with dilutions<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF failed: -> redo today with dilutions<br><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013628') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-06-28</h1><b>Name of participants: </b>Christoffer A, Mikael S, Viktor B, Stephanie H, Viktor T, Jens I, Anders E, Lovisa P., Emil M., Peter C., Ken B.-A., Karl H., Nils A, Magnus B, Pontus D, Alexander W<br><br><h2>Ongoing constructs:</h2><b>L. reuteri:</b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>12. DSM 20016-noplasmid<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-noplasmid<br>11. 36E-noplasmid<br>9. JH2-2 pAMβ1<br><br><b>L. lactis:</b><br>13. MG1363-pJP059<br>15. unknown-pGus<br>16. MG1363-noplasmid<br><br><b>E.coli:</b><br>3. pSB3T5-red<br>4. pSB1A3-red<br>5. pSB4K15<br>7. pSB3K3-red<br>11. pSB4A15<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>19.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>20.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>21.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>23.pSB1C3-CrtB<br>24.pSB1C3-CrtI<br>25.pSB1C3-CrtY<br>42. D5α with plasmid pSB4S15-red <br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>57.B0034-Idi<br>ispA<br><br><h2>Todays work</h2><b>Frozen stock:</b><br>Lb13. Lc. lactis MG1363 pJP059 clone 2<br>Lb15. Lc. lactis unknown pGus clone 2<br>Lb16. Lc. lactis MG1363 noplasmid clone 2<br><br><b>Digest:</b><br>25.pSB1C3-CrtY<br><br><b>Transformation:</b><br>24.pSB1C3-CrtI<br>23.pSB1C3-CrtB<br><br><b>Plasmid preparation:</b><br>Lb13. Lc. lactis MG1363 pJP059 clone 2 “I, II, III(2x1.5 ml culture)”<br>Lb15. Lc. lactis MG1363 pGus clone 2 “I, II, III(2x1.5 ml culture)”<br>3. pSB3T5-red<br>4. pSB1A3-red<br>5. pSB4K15<br>7. pSB3K3-red<br>11. pSB4A15<br><br><b>Re-streaks:</b><br> Lb3. reuteri DSM20016 pVS2<br>Lb8. reuteri 100-23 pGT232*<br>Lb11. plantarum 36E<br>Lb13. Lc. lactis MG1363 pJP059<br>Lb16. Lc. lactis MG1363 noplasmid<br>*from frozen stock<br><br><b>Streaking of strains:</b><br> Lb15. Lc. lactis MG1363 plasmid unknown pGus:<br>Lb18. Lb. delbrueckii bulgaricus (from freeze dried yoghurt culture) <br>Lb19. Lc. Thermophilus (from freeze dried yoghurt culture) <br><br><b>O/N of restreak from Erik Gullberg:</b><br>42. pSB4S15-red, clones 1 and 2<br><br><b>PCR:</b><br>57.B0034-Idi*<br>ispA*<br>19.1**<br>20.1**<br>21.1**<br>* Amplified from D5α-chromosome. <br>** Now with DMSO<br><br><b>Gel electrophoresis:</b><br>57.B0034-Idi<br>ispA<br>19.1<br>20.1<br>21.1<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br><br><b>O/N Screening:</b><br>19.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>20.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>21.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br><br><h2>Results</h2><b>Previous day:</b><br><b>Plasmid preparation:</b><br>Lb13. Lc. lactis MG1363 pJP059 clone 1: 6.7 ng/µl (run 4)<br>→ Store in freezer, try again with new protocol <br><br><b>Transformation:</b><br> Lb10. plantarum 256 pLUL364: No growth<br>Lb11. plantarum 36E pVS2: No growth<br>Lb12. Lc. lactis DSM20016-pVS2: No growth<br>→ Try again later with new competent cells<br><br><b>Wednesday 26/6:</b><br><b>Streaking of strains:</b><br> Lb13. Lc. lactis MG1363 pJP059: Good growth on all concentrations (1x, 5x & 10x) → Re-streak today<br>Lb15. Lc. lactis MG1363 pGus: No growth → Try again on new plates, possibly a problem with antibiotics<br>Lb16. Lc. lactis MG1363 no plasmid: Good growth on all concentrations (1x, 5x & 10x) → Re-streak today<br><br><b>Today:</b><br> <b>Plasmid preparation:</b><br>Lb13. Lc. lactis MG1363-pJP059 clone 2 “I”: 15, 8 ng/µl<br>Lb13. Lc. lactis MG1363-pJP059 clone 2 “II”: 7,5 ng/µl<br>Lb13. Lc. lactis MG1363-pJP059 clone 2 “III(2x1.5 ml culture)”: 48,3 ng/µl <br>Lb15. Lc. lactis unknown-pGus clone 2 “I”: 7,3 ng/µ<br>Lb15. Lc. lactis pGus clone 2 “II”: 8,1 ng/µl<br>Lb15. Lc. lactis pGus clone 2 “III(2x1.5 ml culture)”: 48,1 ng/µl<br>→ Mark and store in freezer.<br><br><b>Gel electrophoresis:</b><br>57.B0034-Idi: hard do decipher results, redo tomorrow <br>ispA: (“)<br>19.1: failed<br>20.1: failed<br>21.1: failed<br>43. pSB1C3-B0034-TAL: failed (image 1 2013.06.28)<br>44. pSB1C3-B0034-His-TAL: failed (“)<br>45. pSB1C3-B0034-4CL: failed (“)<br>46. pSB1C3-B0034-His-4CL: failed (“)<br>47. pSB1C3-B0034-STS: failed (image 2 2013.06.28)<br>48. pSB1C3-B0034-His-STS: failed (“)<br><br><h2>Other experiments</h2><b>THMS buffer preparation</b><br><b>Aliquoting of sigma water</b><br><b>Preparation of antibiotic-free LB-agar plates</b><br><b>Dilution of promotor-oligonucleotides from stock to 5 µM:</b><br>CP1_F<br>CP1_R<br>CP11_F<br>CP11_R <br>CP8_F<br>CP8_R<br>CP44_F<br>CP44_R<br>CP30_F <br>CP30_R <br>CP29_F<br>CP29_R<br>CP41_F<br>CP41_R<br><br><b>Dilution of promotor oligonucleotide from pellet to 100 µM:</b><br>usp45_mut_F<br><br><b>Phosphorylation of promotor-oligonucleotides:</b><br>CP1_F <br>CP1_R <br>CP11_F <br>CP11_R<br>CP8_F<br>CP8_R<br>CP44_F<br>CP44_R<br>CP30_F<br>CP30_R<br>CP29_F<br>CP29_R<br>CP41_F<br>CP41_R</div>'; | ||
| + | } | ||
| + | else if(id == 'd2013627') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-06-27</h1><br><b>Name of participants: </b>Marcus H, Lovisa P, Emil M, Peter C, Ken B-A, Karl H, Kristoffer L, Christoffer A, Mikael S, Jens I, Nafisa B, Stephanie H, Alona N, Viktor T, Viktor B, Nils, Alexander, Thorsten, Pontus, Magnus, Malin<br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri: </b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>12. DSM 20016-noplasmid<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-noplasmid<br>11. 36E-noplasmid<br>9. JH2-2 pAMβ1<br><br><b>L. lactis:</b><br>13. MG1363-pJP059<br>15. unknown-pGus<br>16. MG1363-noplasmid<br><br><b>E.coli:</b><br>3. pSB3T5-red<br>4. pSB1A3-red<br>5. pSB4K15<br>7. pSB3K3-red<br>11. pSB4A15<br>14. pET13b-zifz:4cl-PBSII:STS<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>19.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>20.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>21.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>23.pSB1C3-CrtB<br>24.pSB1C3-CrtI<br>25.pSB1C3-CrtY<br>42. D5α with plasmid pSB4S15-red <br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br><br><h2>Todays work</h2><br><b>Transformation:</b><br>Lb11. plantarum 36E pVS2<br>Lb10. plantarum 256 pLUL634<br>Lb12. plantarum DSM20016 pVS2<br><br><b>Assembly:</b><br>23.Crt.B + pEL3A15<br>24.Crt.I + pEL3K16<br><br><b>Frozen stock: </b><br>Lb9. faecalis JH2-2 pAMβ1<br>Lb13. Lc. lactis MG1363 pJP059 clone 1<br>Lb15. Lc. lactis MG1363 pGus clone 1<br>Lb16. Lc. lactis MG1363 no plasmid clone 1<br><br><b>Plasmid preparation: </b><br>Lb13. Lc. lactis MG1363 pJP059 clone 1 <br>Lb15. Lc. lactis MG1363 pGus clone 1<br>Lb9. faecalis JH2-2 pAMβ1<br><br><b>Over night:</b><br>3. pSB3T5-red<br>4. pSB1A3-red<br>5. pSB4K15<br>7. pSB3K3-red<br>11. pSB4A15<br><br><b>Gel electrophoresis:</b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>19. (digest from previous day)<br>20. (“)<br>21. (“)<br><br><b>PCR amplification of construct:</b><br>miraculin<br>19.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>20.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>21.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>25. pSB1C3-CrtY<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br><br><b>O/N screening PCR:</b><br>19.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>20.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>21.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>25.pSB1C3-CrtY<br><br><h2>Results</h2><br><b>Previous day:</b><br><b>O/N:</b> <br>Lb13. Lc. lactis MG1363 pJP059 clone 2: Visual pellet, foggy medium; nc ok! * <br>Lb15. Lc. lactis MG1363 pGus clone 2: Visual pellet, foggy medium; nc ok! *<br>Lb16. Lc. lactis MG1363 no plasmid clone 2: Visual pellet, foggy medium; nc ok!*<br>Lb13. Lc. lactis MG1363 pJP059 clone 1: Visual pellet, slightly foggy medium; nc ok! **<br>Lb15. Lc. lactis MG1363 pGus clone 1: Visual pellet, slightly foggy medium; nc ok! **<br>Lb16. Lc. lactis MG1363 no plasmid clone 1: Visual pellet, slightly foggy medium; nc ok! **<br>* Optimized conditions = 10µg/ml antibiotics, fill almost up the falcon tube with M17-broth to get rid of the air, let grow in 30o C, make sure that the bacteria pellet is dissolved. → Let grow for one more day <br>** → Frozen stock and plasmid prep.<br><br><b>Transformation: </b><br>42. pSB4S15-red: Many small white colonies, contamination with cocci?<br>→ Discard, get clones from Erik G.<br><br><b>Streaking of strains:</b><br>Lb13. Lc. lactis MG1363 pJP059: Some growth.<br>Lb15. Lc. lactis MG1363 pGus: Hard to see without opening lock.<br>Lb16. Lc. lactis MG1363 no plasmid: Hard to see without opening lock.<br>→ let grow one more day.<br><br><b>Today:</b><br><b>Plasmid preparation: </b><br>Lb9. faecalis JH2-2 pAMβ1 clone 1: 10.1 ng/µl<br>Lb9. faecalis JH2-2 pAMβ1 clone 2: 24.7 ng/µl<br>Lb13. Lc. lactis MG1363-pJP059 clone 1: 4.9 ng/µl (run 1)<br>Lb13. Lc. lactis MG1363-pJP059 clone 1: 6.0 ng/µl (run 2)<br>Lb13. Lc. lactis MG1363-pJP059 clone 1: 6.1 ng/µl (run 3)<br>Lb15. Lc. lactis MG1363 pGus clone 1: 61 ng/µl (run 1)<br>→ Measure again tomorrow<br><br><b>Gel electrophoresis:</b><br>Miraculin: fail, problems with primers, probably due to problematic secondary structure.<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT: failed, ladder too, probably problem with gel. Image 1 (2013.06.27)<br>19. (digest from previous day): Success<br>20. (“): Success<br>21. (“): Success<br>19. (PCR): Fail -> redo with lower TM<br>20. (“): Fail -> redo with lower TM<br>21. (“): Fail -> redo with lower TM<br>25. pSB1C3-CrtY: Good results<br><br><h2>Other experiments</h2><br><b>Preparation of THMS buffer</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013626') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h2>Wednesday 2013-06-26</h2><br><b>Name of participants: </b>Christoffer A, Stephanie H, Alona N, Anders E, Mikael S, Anton B, Viktor B, Marcus H, Lovisa P, Emil M, Ken B-A, Karl H, Theodor L, Magnus B., Malin B., Thorsteinn O., Nils A., Alexander W., Niclas S., Pontus D., Christoffer F.<br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri: </b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>12. DSM 20016-noplasmid<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-noplasmid<br>11. 36E-noplasmid<br><br><b>E. faecalis:</b><br>9. JH2-2 pAMβ1<br><br><b>L. lactis</b><br>13. MG1363-pJP059<br>15. unknown-pGus<br>16. MG1363-noplasmid<br><br><b>E.coli:</b><br>14. pET13b-zifz:4cl-PBSII:STS<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>25.pSB1C3-CrtY<br>19.psB4C5-CrtE~O-Z / w. linker, ZF & RBS<br>20.psB4C5-CrtE~Z / w. linker, ZF & RBS<br>21.psB4C5-CrtE~I / w. linker, ZF & RBS<br>42. D5α with plasmid pSB4S15-red<br><br><h2>Todays work</h2><br><b>Competent cells preparation:</b><br>Lb11. plantarum 36E no plasmid<br>Lb10. plantarum 256 no plasmid<br>Lb12. reuteri DSM 20016 no plasmid<br><br><b>Transformation:</b><br>plantarum 36E pAMB1<br>plantarum 256 pAMB1<br>Lb3. reuteri DSM 20016 pVS2<br>25.pSB1C3-CrtY<br>42. pSB4S15-red <br><br><b>O/N:</b><br>Lb5.2 plantarum 256 pAMβ1*<br>Lb8.2 reuteri 100-23 pGT232*<br>Lb13. Lc. lactis MG1363 pJP059**<br>Lb15. Lc. lactis MG1363 pGus**<br>Lb16. Lc. lactis MG1363 no plasmid**<br>*Two tubes of 8.2 were prepared. It has grown rather poorly to not at all before. If it continues to grow poorly we may have to redo O/N from a plate rather than from our frozen stock.<br>** Optimized conditions → 10µg/ml antibiotics, fill almost up the falcon tube with M17-broth to get rid of the air, let grow in 30o C, make sure that the bacteria pellet is dissolved. <br><br><b>Digestion:</b><br>19.1.psB4C5-CrtE~O-Z / w. linker, ZF & RBS<br>19.2psB4C5-CrtE~O-Z / w. linker, ZF & RBS<br>20.1.psB4C5-CrtE~Z / w. linker, ZF & RBS<br>20.2.psB4C5-CrtE~Z / w. linker, ZF & RBS<br>21.1.psB4C5-CrtE~I / w. linker, ZF & RBS<br>21.2.psB4C5-CrtE~I / w. linker, ZF & RBS<br><br><b>Gel electrophoresis:</b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br><br><b>PCR amplification of construct:</b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT (third try, using same concentration of template as second try but lower annealing temperature )<br>19.1.psB4C5-CrtE~O-Z / w. linker, ZF & RBS<br>19.2psB4C5-CrtE~O-Z / w. linker, ZF & RBS<br>20.1.psB4C5-CrtE~Z / w. linker, ZF & RBS<br>20.2.psB4C5-CrtE~Z / w. linker, ZF & RBS<br>21.1.psB4C5-CrtE~I / w. linker, ZF & RBS<br>21.2.psB4C5-CrtE~I / w. linker, ZF & RBS<br><br><br><h2>Results</h2><br><b>Previous day:</b><br><b>O/N:</b> <br>Lb13. Lc. lactis MG1363 pJP059: Growing, visual pellet.*<br>Lb15. Lc. lactis MG1363 pGus: Growing, visual pellet.*<br>Lb16. Lc. lactis MG1363 no plasmid: Growing, visual pellet.*<br>*Weak growth in the morning, better in the afternoon. → Let grow for one more day + new O/N with optimized conditions.<br><br><b>Streaking of strains: </b><br>Lb13. Lc. lactis MG1363 pJP059: No visual growth → Let grow for one more day.<br>Lb15. Lc. lactis MG1363 pGus: No visual growth → --”--<br>Lb16. Lc. lactis MG1363 no plasmid: No visual growth → --”—<br><br><b>Today:</b><br><b>Gel electrophoresis:</b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT: Failed, possibly nonspecific primer binding, not enough template DNA. Image 1 (2013.06.26)<br><br><b>Transformation:</b><br>25.pSB1C3-CrtY: failed<br>D5alpha competent cells were competent<br><br><br><h2>Other experiments</h2><br><b>Preparation of cold electroporation buffer for competent cells</b><br><br><b>Preparation of 50% glycerol stock: </b><br>500 ml 50% glycerol stock was prepared from 86-88% glycerol.</div>' | ||
| + | } | ||
| + | else if(id == 'd2013625') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-06-25</h1><br><br><b>Name of participants:</b> Marcus H, Lovisa P, Emil M, Ken B-A, Karl H, Peter C, Theodor L, Christoffer A, Nafisa B, Stephanie H, Alona N, Anders E, Viktor T, Viktor B, Mikael S, Anton B<br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri: </b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>12. DSM 20016-noplasmid<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-noplasmid<br>11. 36E-noplasmid<br><br><b>E. faecalis:</b><br>9. JH2-2 pAMβ1<br><br><b>L. lactis:</b><br>13. MG1363-pJP059<br>15. unknown-pGus<br>16. MG1363-noplasmid<br>14. pET13b-zifz:4cl-PBSII:STS<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br><br><h2>Todays work</h2><br><b>Gel electrophoresis:</b><br>14. pET13b-zifz:4cl-PBSII:STS<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br><br><b>PCR amplification of construct:</b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT (second try, using smaller concentration of template and higher annealing temperature )<br>Lb13. Lc. lactis MG1363 pJP059<br><br><b>Competent cells preparation: </b><br>Lb11. plantarum 36E no plasmid<br>Lb10. plantarum 256 noplasmid<br>Lb12. reuteri DSM 20016 no plasmid<br><br><b>Plasmid preparation: </b><br>Lb6.1 reuteri DSM 20016 pLUL634TsA8<br>Lb6.2 reuteri DSM 20016 pLUL634TsA8<br>Lb6.3 reuteri DSM 20016 pLUL634TsA8<br>Lb6.4 reuteri DSM 20016 pLUL634TsA8 *<br>Lb7.2 reuteri DSM 20016 pGFP<br>Lb7.3 reuteri DSM 20016 pGFP<br>Lb7.4 reuteri DSM 20016 pGFP *<br>Lb14.1 reuteri DSM 20016 pLUL631<br>Lb14.2 reuteri DSM 20016 pLUL631<br>Lb14.3 reuteri DSM 20016 pLUL631<br>Lb14.4 reuteri DSM 20016 pLUL631 *<br>* 6.4, 7.4 and 14.4 were treated in a different way than the regular plasmid prep protocol for L. reuteri. Instead of treating them with LYSIS-buffer and lysozyme they were treated with resuspension solution and lysis solution from the plasmid prep kit and allowed to lyse for ~30 minutes. The rest were treated as usual, but with 1 ml of 10 mg/ml lysozyme instead of 2 ml.<br><br><b>O/N:</b> <br>Lb13. Lc. lactis MG1363 pJP059<br>Lb15. Lc. lactis MG1363 pGus<br>Lb16. Lc. lactis MG1363 no plasmid<br><br><b>Streaking of strains:</b><br>Lb13. Lc. lactis MG1363 pJP059<br>Lb15. Lc. lactis MG1363 pGus<br>Lb16. Lc. lactis MG1363 no plasmid<br><br><b>PCR amplification: </b><br>Lb13. Lc. lactis MG1363 pJP059*<br>*Another attempt at colony PCR with different concentrations of lysozyme and buffer.<br><br><h2>Results</h2><br><b>Gel electrophoresis:</b><br>15. failed. Possibly nonspecific primer binding, too much template DNA. Image 1 (2013.06.25)<br>Lb13. Lc. lactis MG1363 pJP059: Only short bands, probably primer dimers because lysing failed. → Try again with a plasmid prep instead of culture.<br><br><b>Competant cells:</b><br>Wrong volume was added. → Try again tomorrow with the right volume.<br><br><b>Plasmid preparation: *</b><br>Lb6.1 reuteri DSM 20016 pLUL634 TsA8: 203.7 ng/µl<br>Lb6.2 reuteri DSM 20016 pLUL634 TsA8: 203.2 ng/µl<br>Lb6.3 reuteri DSM 20016 pLUL634 TsA8: 154.8 ng/µl<br>Lb6.4 reuteri DSM 20016 pLUL634 TsA8: 39.5 ng/µl<br>Lb7.2 reuteri DSM 20016 pGFP: 100.5 ng/µl<br>Lb7.3 reuteri DSM 20016 pGFP: 141.2 ng/µl<br>Lb7.4 reuteri DSM 20016 pGFP: 8.7 ng/µl<br>Lb14.1 reuteri DSM 20016 pLUL631: 162.7 ng/µl<br>Lb14.2 reuteri DSM 20016 pLUL631: 85.2 ng/µl<br>Lb14.3 reuteri DSM 20016 pLUL631: 49.8 ng/µl<br>Lb14.4 reuteri DSM 20016 pLUL631: 7.7 ng/µl<br>*The alternative way of lysing the cells proved to be quite ineffective so we will continue to use our standard protocol. The earlier plasmid preparations of the above “constructs”, that had far poorer concentrations, were replaced with these better ones.<br><br><h2>Other experiments</h2><br><b>Preparation of MRS broth </b><br><br><b>Making plates for Lc. lactis cultivation (from M17-broth)</b><br><br><b>Preparation of homemade yogurt from dried bacteria and from commercial yogurt with active bacteria culture</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013624') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-06-24</h1><br><b>Name of participants: </b>Christoffer A, Nafisa B, Stephanie H, Alona N, Anders E, Viktor T, Viktor B, Mikael S, Anton B, Marcus H, Lovisa P, Emil M, Ken B-A, Karl H, Peter C, Alexander W., Malin B., Magnus B., Thorsteinn O., Niclas S., Pontus D.<br><br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri: </b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>12. DSM 20016-noplasmid<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-noplasmid<br>11. 36E-noplasmid<br><br><b>E. faecalis:</b><br>9. JH2-2 pAMβ1<br><br><b>L. lactis</b><br>13. MG1363-pJP059<br><br><b>E. coli:</b><br>14. pET13b-zifz:4cl-PBSII:STS<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m <br><br><h2>Todays work</h2><br><b>Plasmid preparation:</b><br>Lb8. reuteri 100-23 pGT232<br>Lb9. faecalis JH2-2 pAMβ1<br><br><b>Digestion of construct:</b><br>14. pET13b-zifz:4cl-PBSII:STS<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br><br><b>O/N (from frozen stock): </b><br>Lb2. 1063 pLUL631<br>Lb6. DSM 20016 pLUL63TsA8<br>Lb7. DSM 20016 pGFP<br>Lb8. 100-23 pGT232<br>Lb9. faecalis JH2-2 pAMβ1<br><br><b>PCR:</b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT*<br>Lb13. Lc. lactis MG1363 pJP059*<br>*Colony-PCR. Cells might need lysing. Two samples from the same stock was resuspended in 100μl distilled water. Lyzosyme and buffer was added to one of the samples followed by heating to 95 degrees celsius. <br><br><b>Transformation test of competent cells (D5alpha strain number 10) with plasmid preps: </b><br>1.1. pSB1C3-red<br>18.2. pSB1C3-CHS_m<br>40. pEL3A17-red<br><br><h2>Results</h2><br><b>Plasmid preparation:</b><br>Lb8. Lc. lactis 100-23 pGT232 clone 2: 14.8 ng/μl<br>Lb9. Lc. lactis JH2-2 pAMβ1 clone 1: 13.5 ng/μl<br>Lb9. Lc. lactis JH2-2 pAMβ1 clone 2: 13.4 ng/μl<br>Lb9. faecaliJH2-2 pAMβ1 clone 4: 0.5 ng/μl<br>Plasmid concentrations were low on the plasmid preparations from today and the ones done on 22/6-13. New plasmid preparations will be done tomorrow on these strains from the O/N that were done today.<br><br><h2>Other experiments</h2><br><b>Primer stock preparation</b><br><br><b>Dilution of sequencing primers: </b><br>3 tubes of 5 µM VF2-primer, 200 uL<br>3 tubes 5 µM VR-primer, 200 µL<br><br><b>Re-measuring concentration of earlier plasmid preps: *</b><br>1.4 CF48-pLR581: 127.8 ng/µl<br>1.5 CF48-pLR581: 215.9 ng/µl<br>3.2 DSM 20016-pVs2: 201.2 ng/µl<br>3.3 DSM 20016-pVs2: 139.1 ng/µl<br>3.10 DSM 20016-pVs2: 183.5 ng/µl<br>*We deduced that the low, and sometimes even negative, concentrations were caused by a bubble when loading the samples. Changes to protocol have been made with this in mind.<br><br><b>Preparing M17-broth, 950 ml.</b><br><b>(“) SOB</b><br><b>(“) CCMB80 (20% glycerol, pH 6,8)</b><br><b>Alliquoting dNTP, buffer, enzymes</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013623') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Sunday 2013-06-23</h1><br><b>Name of participants: </b>Hampus E, Alexander W<br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri: </b><br>4. 100-23-noplasmid<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br><br><b>E. coli: </b><br>23.pCrcSB1C3-Crt-B<br>24.pSB1C3-Crt-I<br>25. pSB1C3-CrtY<br><br><h2>Todays work</h2><br><b>Glycerol stock preparation:</b><br>Lb4. reuteri 100-23 no plasmid<br>Lb5. plantarum 256 rifR-Pamβ1<br>23.pCrcSB1C3-Crc-B<br>24.pSB1C3-Crc-I<br><br><b>Restreak:</b><br>25. pSB1C3- crtY<br> <br><b>Plasmidpreparation </b><br>23.pCrcSB1C3-Crc-B<br>24.pSB1C3-Crc-I<br><br><h2>Result</h2><br><b>O/N:</b><br>25.pSB1C3-crtY: One of the overnights did not grow<br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013622') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-06-22</h1><br><b>Name of participants: </b>Nafisa B, Mikael S, Marcus H<br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri: </b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>12. DSM 20016-noplasmid<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-noplasmid<br>11. 36E-noplasmid<br><br><b>E. faecalis:</b><br>9. JH2-2 pAMβ1<br><br><b>E. coli:</b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>6. pSB1K3-red<br>14. pET13b-zifz:4cl-PBSII:STS<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br><br><h2>Todays work</h2><br><b>Glycerol stock preparation: </b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>6. pSB1K3-red <br>Lb6. reuteri DSM 20016-pLUL63TsA8<br>Lb7. reuteri DSM 20016-pGFP<br>Lb8. reuteri 100-23-pGT232<br>Lb9. faecalis JH2-2- pAMβ1<br>Lb10. plantarum 256-noplasmid<br>Lb11. plantarum 36E-noplasmid<br>Lb12. reuteri DSM 20016-noplasmid<br>Lb14. reuteri DSM 20016-pLUL631 (B?)<br><br><b>O/N: </b><br>Lb4. reuteri100-23-noplasmid<br>Lb5. plantarum 256-rifR-pAMβ1<br><br><b>Plasmid preparation:</b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>6. pSB1K3-red <br>Lb14. DSM 20016-pLUL631 (B?)<br>Lb6. DSM 20016-pLUL63TsA8<br>Lb7. DSM 20016-pGFP<br><br><h2>Results</h2><br><b>Plasmid preparation:</b><br><b>L. reuteri:</b><br>Lb14.1 (strain 14, clone 1) DSM 20016-pLUL631 (B?): 10.0 ng/μl<br>Lb14.2 DSM 20016-pLUL631 (B?): 13.6 ng/μl<br>Lb14.3 DSM 20016-pLUL631 (B?): 14.7 ng/μl<br>Lb14.4 DSM 20016-pLUL631 (B?): 28,4 ng/μl<br>Lb6.1 DSM 20016-pLUL63TsA8: 18.7 ng/μl<br>Lb6.2 DSM 20016-pLUL63TsA8: 14.4 ng/μl<br>Lb6.3 DSM 20016-pLUL63TsA8: 12.5 ng/μl<br>Lb6.4 DSM 20016-pLUL63TsA8: 27.7 ng/μl<br>Lb7.2 DSM 20016-pGFP: 29.9 ng/μl<br>Lb7.3 DSM 20016-pGFP: 16.9 ng/μl<br>Lb7.4 DSM 20016-pGFP: 10.9 ng/μl</div>' | ||
| + | } | ||
| + | else if(id == 'd2013621') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-06-21</h1><br><b>Name of participants: </b>Stephanie H, Alona N, Anton B, Mikael S, Nafisa B, Anders E, Marcus H, Ken B-A, <br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri: </b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>12. DSM 20016-noplasmid<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-noplasmid<br>11. 36E-noplasmid<br><br><b>E. faecalis:</b><br>9. JH2-2 pAMβ1<br><br><b>E. coli:</b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>6. pSB1K3-red<br>14. pET13b-zifz:4cl-PBSII:STS<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>22. pSB1C3-B0032-BFP<br><br><h2>Todays work</h2><br><b>Plasmid preparation*: </b><br>Lb1. reuteri CF48-pLR581<br>Lb3. reuteri DSM 20016-pVs2 <br><i>*to get more useable material and to look for different result from clones grown in a lock and a CO2-cupboard.</i><br><br><b>O/N: </b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>6. pSB1K3-red<br>Lb6. reuteri DSM 20016-pLUL63TsA8<br>Lb7. reuteri DSM 20016-pGFP<br>Lb14. reuteri DSM 20016-pLUL631(B?)<br>Lb12. reuteri DSM 20016-noplasmid<br>Lb8. reuteri 100-23-pGT232<br>Lb10. plantarum 256-noplasmid<br>Lb11. plantarum 36E-noplasmid<br>Lb9. faecalis JH2-2 pAMβ1 *-<br><i>*from the plate without erythromycin. </i><br><br><b>Plasmid preparation:</b><br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>22. pSB1C3-B0032-BFP<br><br><h2>Results</h2><br><b>Previous day:</b><br><b>Re-streak: </b><br>Lb4. reuteri 100-23-noplasmid: No growth. → Grow on non-antibiotic plates <br><br><b>Today:</b><br><b>Plasmid preparation:</b><br>Lb1.2 (strain 1, clone 2) reuteri CF48-pLR 581: Non-usable concentration *<br>Lb1.4 reuteri CF48-pLR 581: 161.8 ng/µl *<br>Lb3.8 reuteri DSM 20016 pVS2: 148.5 ng/µl *<br>Lb3.10 reuteri DSM 20016 pVS2: Non-usable concentration *<br><i>*there is seemingly no correlation between growth in lock vs. CO2-cupboard and plasmid concentration. This seems strange though. Cupboard is not supposed to create an anaerobic environment → do not use cupboard</i><br><br><h2>Other experiments</h2><br><b>SET buffer preparation: </b><br><b>Lysis buffer preparation: </b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013620') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-06-20</h1><br><b>Name of participants: </b>Christoffer A, Stephanie H, Alona N, Viktor T, Viktor B, Mikael S, Anton B, Anders E, Emil M, Marcus H, Ken B-A, Lovisa P, Peter C, Malin B., Pontus D., Magnus B., Alexander W., Niclas S., Christoffer F.<br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri: </b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>12. DSM 20016-noplasmid<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-noplasmid<br>11. 36E-noplasmid<br><br><b>E. faecalis:</b><br>9. JH2-2 pAMβ1<br><br><b>E. coli:</b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>6. pSB1K3-red<br>14. pET13b-zifz:4cl-PBSII:STS<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>22. pSB1C3-B0032-BFP<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br>23. pSB1C3-CrtB<br>24. pSB1C3-CrtI<br>26. pEL3K15-red<br>27. pEL3K16-red<br>28. pEL3K17-red<br>29. pEL3K17(FRT)-red<br>30. pEL3C15-red<br>31. pEL3C16-red<br>32. pEL3C17-red<br>33. pEL3C17(FRT)-red<br>34. pEL3S15-red<br>35. pEL3S16-red<br>36. pEL3S17-red<br>37. pEL3S17(FRT)-red<br>38. pEL3A15-red<br>39. pEL3A16-red<br>40. pEL3A17-red<br>41. pEL3A17(FRT)-red<br><br><br><h2>Todays work:</h2><br><b>Glycerol stock preparation: </b><br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br>Lb1. reuteri CF48-pLR581<br>Lb2. reuteri 1063-pLUL631<br>Lb3. reuteri DSM 20016-pVS2 <br><br><b>Plasmid preparation:</b><br>14. pET13b-zifz:4cl-PBSII:STS<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br>22. pSB1C3-B0032-BFP<br>26. pEL3K15-red<br>26. pEL3K15-red<br>27. pEL3K16-red<br>28. pEL3K17-red<br>29. pEL3K17(FRT)-red<br>30. pEL3C15-red<br>31. pEL3C16-red<br>32. pEL3C17-red<br>33. pEL3C17(FRT)-red<br>34. pEL3S15-red<br>35. pEL3S16-red<br>36. pEL3S17-red<br>37. pEL3S17(FRT)-red<br>38. pEL3A15-red<br>39. pEL3A16-red<br>40. pEL3A17-red<br>41. pEL3A17(FRT)-red<br>Lb1. reuteri CF48-pLR581<br>Lb2. reuteri 1063-pLUL631<br>Lb3. reuteri DSM 20016-pVS2 <br><br><b>O/N: </b><br>Lb2. reuteri 1063-pLUL631<br>Lb1. reuteri CF48-pLR581<br>Lb3. reuteri DSM 20016-pVs2 <br><br><b>Streaking of new strains from Stefan Roos: </b><br>Lb6. reuteri DSM 20016-pLUL63TsA8<br>Lb7. reuteri DSM 20016-pGFP<br>Lb12. reuteri DSM 20016-noplasmid *<br>Lb14. reuteri DSM 20016-pLUL631(B?)<br>Lb4. reuteri 100-23-noplasmid<br>Lb8. reuteri 100-23-pGT232 *<br>Lb5. plantarum 256-rifR-pAMβ1<br>Lb10. plantarum 256-noplasmid *<br>Lb11. plantarum 36E-noplasmid *<br>Lb9. faecalis JH2-2 pAMβ1 **<br><i>*both on burned broth without glucose and CaCl2 and regular one.</i><br><i>**re-streaked on erythromycin and on antibiotic-free plates.</i><br><br><b>Transformation:</b><br>23. pSB1C3-CrtB<br>24. pSB1C3-CrtI<br><br><b>Restreaks:</b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>6. pSB1K3-red<br><br><h2>Results</h2><br><b>Previous day:</b><br>Lb13. Lc. lactis MG1363-pJP059 : No growth those strain. → Try another medium (e.g. M17) <br><br><b>Today:</b><br><b>Plasmid preparation: </b><br>19.1: 50.5 ng/µl <br>19.2: 33.6 ng/µl<br>19.3: 45.2 ng/µl<br>19.4: 36.5 ng/µl<br>20.1: 28.9 ng/µl<br>20.2: 29.6 ng/µl<br>20.3: 41.8 ng/µl<br>20.4: 35.8 ng/µl<br>21.1: 47.7 ng/µl<br>21.2: 23.2 ng/µl<br>21.3: 38.8 ng/µl<br>21.4: 48.9 ng/µl<br>Lb1. reuteri CF48-pLR 581 clone 1: 74.8 ng/µl<br>Lb1. reuteri CF48-pLR 581 clone 2: Non-usable concentration → try again.<br>Lb1. reuteri CF48-pLR 581 clone 3: 77.5 ng/µl<br>Lb1. reuteri CF48-pLR 581 clone 4: Non-usable concentration → try again.<br>Lb2. reuteri 1063-pLUL 631 clone 1: 70.3 ng/µl<br>Lb2. reuteri 1063-pLUL 631 clone 2: 39.4 ng/µl<br>Lb2. reuteri 1063-pLUL 631 clone 3: 72.4 ng/µl<br>Lb2. reuteri 1063-pLUL 631 clone 6: 61.0 ng/µl<br>Lb3. reuteri DSM 20016 pVS2 clone 1: 231.7 ng/µl<br>Lb3. reuteri DSM 20016 pVS2 clone 2: Non-usable concentration → try again.<br>Lb3. reuteri DSM 20016 pVS2 clone 3: Non-usable concentration → try again.<br>Lb3. reuteri DSM 20016 pVS2 clone 4: 86.3 ng/µl<br><br><br><h2>Other experiments</h2><br><b>Antibiotic-free MRS-broth plates preparation: </b><br><b>SET-buffer and Lysis-buffer preparation: </b><br><b>MRS-broth agar preparation: </b><br><b>Preparation of plates with and without antibiotic:</b> </div>' | ||
| + | } | ||
| + | else if(id == 'd2013619') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-06-19</h1><br><br><b>Name of participants: </b>Emil M, Marcus H, Ken B-A, Sabri J., Christoffer A, Nafisa B, Stephanie H, Alona N, Anders E, Jens I, Viktor T, Viktor B, Mikael S, Anton B, Malin B, Pontus D, Magnus B, Alexander W, Thorsteinn O, Niclas S, Nils A<br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri: </b><br>2. 1063-pLUL631<br>1. CF48-pLR581<br>3. DSM20016-pVs2<br><br><b>Lactoccocus lactis:</b><br>13. MG1363-pJP059<br><br><b>E. coli:</b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>3. pSB3T5-red<br>4. pSB1A3-red<br>6. pSB1K3-red<br>7. pSB3K3-red<br>12. pSB4K5-red<br>14. pET13b-zifz:4cl-PBSII:STS<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br>22. pSB1C3-B0032-BFP<br>23. pSB1C3-CrtB<br>24. pSB1C3-CrtI<br>25. pSB1C3-CrtY<br>26. pEL3K15-red<br>27. pEL3K16-red<br>28. pEL3K17-red<br>29. pEL3K17(FRT)-red<br>30. pEL3C15-red<br>31. pEL3C16-red<br>32. pEL3C17-red<br>33. pEL3C17(FRT)-red<br>34. pEL3S15-red<br>35. pEL3S16-red<br>36. pEL3S17-red<br>37. pEL3S17(FRT)-red<br>38. pEL3A15-red<br>39. pEL3A16-red<br>40. pEL3A17-red<br>41. pEL3A17(FRT)-red <br><br><h2>Todays work</h2><br><b>Re-streak:</b><br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>22. pSB1C3-B0032-BFP<br>Lb1. reuteri CF48-pLR581<br>Lb2. reuteri 1063-pLUL631<br>Lb3. reuteri DSM20016-pVs2<br><br><b>Spreading:</b> <br>Lb13. Lc. lactis MG1363-pJP059 <br><br><b>O/N:</b> <br>14. pET13b-zifz:4cl-PBSII:STS<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br>26. pEL3K15-red<br>27. pEL3K16-red<br>28. pEL3K17-red<br>29. pEL3K17(FRT)-red<br>30. pEL3C15-red<br>31. pEL3C16-red<br>32. pEL3C17-red<br>33. pEL3C17(FRT)-red<br>34. pEL3S15-red<br>35. pEL3S16-red<br>36. pEL3S17-red<br>37. pEL3S17(FRT)-red<br>38. pEL3A15-red<br>39. pEL3A16-red<br>40. pEL3A17-red<br>41. pEL3A17(FRT)-red<br>Lb1. reuteri CF48-pLR581<br>Lb2. reuteri 1063-pLUL631<br>Lb3. reuteri DSM20016-pVs2<br><br><b>Transformation:</b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>3. pSB3T5-red<br>4. pSB1A3-red<br>6. pSB1K3-red<br>7. pSB3K3-red<br>12. pSB4K5-red<br>23. pSB1C3-CrtB<br>24. pSB1C3-CrtI<br>25. pSB1C3-CrtY<br><br><h2>Results</h2><br><b>Previous day: </b><br><b>Spreading:</b><br>Lb13. Lc. lactis MG1363-pJP059 : No growth. → Try growing with less oxygen. <br><br><b>Transformation</b><br>1. pSB1C3-red: Successful<br>2. pSB3C5-red: Successful<br>3. pSB3T5-red: fail<br>4. pSB1A3-red: fail<br>6. pSB1K3-red: Successful<br>7. pSB3K3-red: fail<br>12. pSB4K5-red: fail<br><br><b>Restreak:</b><br>16. pSB1C3-TAL_m: Successful<br>17. pSB1C3-4Cl_m: Successful<br>18. pSB1C3-CHS_m: Successful<br>22. pSB1C3-B0032-BFP: Successful<br><br><b>Today:</b><br><b>O/N: </b><br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF: Growth<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF: Growth<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF: Growth<br><br><b>Transformation:</b><br>23. pSB1C3-CrtB: fail<br>24. pSB1C3-CrtI: fail<br>25. pSB1C3-CrtY: fail<br><br><h2>Other experiments</h2><br><b>LB-agar plates </b><br><b>CaCl2-solution</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013618') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-06-18</h1><br><b>Name of participants: </b>Malin B, Pontus D, Magnus B, Alexander V, Thorsteinn O, Niclas S, Nils A, Christoffer F, Kristoffer L, Emil M, Theodor L, Marcus H, Ken B.-A, Peter C, Christoffer A, Nafisa B, Stephanie H, Alona N, Anders E, Jens I, Viktor T, Viktor B, Mikael S, Anton B<br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri: </b><br>2. 1063-pLUL631<br>1. CF48-pLR581<br>3. DSM20016-pVs2<br> <br><b>Lactoccocus lactis:</b><br>13. MG1363-pJP059 <br><br><b>E. coli:</b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>3. pSB3T5-red<br>4. pSB1A3-red<br>6. pSB1K3-red<br>7. pSB3K3-red<br>12. pSB4K5-red<br>14. pET13b-zifz:4cl-PBSII:STS<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br>22. pSB1C3-B0032-BFP<br><br><h2>Todays work:</h2><br><b>Streaking of strains from Stefan Roos: </b><br>Lb1. reuteri CF48-pLR581<br>Lb2. reuteri 1063-pLUL631 <br>Lb3. reuteri DSM 20016-pVS2<br>Lb13. Lc. lactis MG1363-pJP059 <br><br><b>Transformation</b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>3. pSB3T5-red<br>4. pSB1A3-red<br>6. pSB1K3-red<br>7. pSB3K3-red<br>12. pSB4K5-red<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>22. pSB1C3-B0032-BFP<br><br><b>Restreak</b><br>14. pET13b-zifz:4cl-PBSII:STS<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br><br><h2>Results:</h2><br><b>Previous day</b><br><b>Restreak:</b><br>14. pET13b-zifz:4cl-PBSII:STS: Growth<br><br><b>Today</b><br><b>Transformation:</b><br>1. pSB1C3-red: fail<br>2. pSB3C5-red: fail<br>3. pSB3T5-red: fail<br>4. pSB1A3-red: fail<br>6. pSB1K3-red: fail<br>7. pSB3K3-red: fail<br>12. pSB4K5-red: fail<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT: fail<br>16. pSB1C3-TAL_m: successful<br>17. pSB1C3-4Cl_m: successful<br>18. pSB1C3-CHS_m: successful<br>22. pSB1C3-B0032-BFP: successful<br><br><b>Restreak:</b><br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF: Growth<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF: Growth<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF: Growth<br><br><h2>Other experiments:</h2><br><b>LB-agar</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013617') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-06-17</h1><br><b>Name of participants: </b>Kristoffer L, Emil M, Lovisa P, Theodor L, Marcus H, Ken B-A, Peter C, Christoffer A, Anton B, Nafisa B, Stephanie H, Alona N, Anders E, Malin B., Pontus D., Alexander V., Magnus B., Nils A. Thorsteinn O., Christoffer F<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>3. pSB3T5-red<br>4. pSB1A3-red<br>6. pSB1K3-red<br>7. pSB3K3-red<br>12. pSB4K5-red<br>14. pET13b-zifz:4cl-PBSII:STS<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br><br><h2>Todays work:</h2><br><b>Transformation</b><br>1. pSB1C3-red<br>2. pSB3C5-red<br>3. pSB3T5-red<br>4. pSB1A3-red<br>6. pSB1K3-red<br>7. pSB3K3-red<br>12. pSB4K5-red<br>14. pET13b-zifz:4cl-PBSII:STS<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>16. pSB1C3-TAL_m<br>17. pSB1C3-4Cl_m<br>18. pSB1C3-CHS_m<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF<br><br><h2>Results:</h2><br><b>Today:</b><br><b>Transformation:</b><br>1. pSB1C3-red: fail<br>2. pSB3C5-red: fail<br>3. pSB3T5-red: fail<br>4. pSB1A3-red: fail<br>6. pSB1K3-red: fail<br>7. pSB3K3-red: fail<br>12. pSB4K5-red: fail<br>14. pET13b-zifz:4cl-PBSII:STS: successful<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>16. pSB1C3-TAL_m: fail<br>17. pSB1C3-4Cl_m: fail<br>18. pSB1C3-CHS_m: fail<br>19. pSB4C5-CrtE~I /w RBS, Linker & ZF: successful<br>20. pSB4C5-CrtE~Z /w RBS, Linker & ZF: successful<br>21. pSB4C5-CrtE~O-Z /w RBS, Linker & ZF: successful<br><br><h2>Other experiments:</h2><br><b>Mixing MRS-broth</b><br><b>Making of EDTA solution</b><br><b>MRS-plates preparation</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013613') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-06-13</h1><br><b>Name of participants:</b> Alexander W, Magnus B, Malin B, Thorsteinn O, Christoffer F.<br><br><h2>Other experiments</h2><br><b>Making competent D5α</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013612') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-06-12</h1><br><b>Name of participants: </b>Alexander W, Magnus B, Malin B, Thorsteinn O, Christoffer F., Pontus D<br><br><h2>Other experiments</h2><br><b>LB-agar plates</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013610') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-06-10</h1><br><b>Name of participants: </b>Alexander W, Magnus B, Malin B, Thorsteinn O, Nils A, Christoffer F., Pontus D, Niclas S.<br><br><h2>Other experiments:</h2><br><b>LB</b><br><b>CaCl2</b><br><b>Antibiotics</b></div>' | ||
| + | } | ||
| + | else if(id == 'd201373') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-07-03</h1><br><b>Name of participants:</b> Lovisa P, Kristoffer L, Emil M, Marcus H, Karl H, Peter C, Ken B.-A, Nils A, Alexander W, Christoffer, Thorsteinn, Magnus, Christoffer Ahlström (CA), Mikael Strandgren (MS), Viktor Blomkvist (VB), Stephanie Herman (SH), Anton Berglund (AB)<br><br><h2>Ongoing constructs:</h2><br><b>E.coli (strain D5α):</b><br>1. pSB1C3-red<br>7. pSB3K3-red<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>57. pSB1C3-B0034-Idi<br>58. pSB1C3-B0034-ispA<br>42. pSB4S15-red<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>77. pSB4C15-red<br><br><b>L. plantarum:</b><br>Lb10. 256-noplasmid<br>Lb11. 36E-noplasmid<br><br><br><b>L. reuteri:</b><br>Lb12. DSM 20016-noplasmid<br><br><b>L. lactis:</b><br>Lb13. MG1363-pJP059<br><br><h2>Todays work</h2><br><b>PCR:</b><br>Lb13.2.3 Lc. lactis MG1363-pJP059 (plasmid)*<br>Lb13.3 Lc. lactis MG1363-pJP059 (colony)*<br>Lb13.4 Lc. lactis MG1363-pJP059 (colony)*<br><i>*Two PCR tubes of 13.2.3 were prepared along with one of 13.3 and one of 13.4.</i><br>Lb13.2.2 lactis MG1363 pJP059 (plasmid)**<br>Lb13.2.3 lactis MG1363 pJP059 (plasmid)**<br>Lb13.3 lactis MG1363 pJP059 (colony)**<br>Lb13.4 Lc. lactis MG1363 pJP059 (colony)**<br><i>**Gel will be done tomorrow. Followed the PCR protocol with DMSO.</i><br>78. pEL3C18-CrtY*<br><i>-*PCR of CrtZ with lower ™-temp(62 insted of 63.6 which was used last time)</i><br><br>Digest:<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>25. pSB1C3-CrtY<br>27. pEL3K16-red<br>51. pEL3C18-red<br><br><b>Ligation:</b><br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>75. pEL3A15-CrtB<br>76. pEL3K16-CrtI<br>78. pEL3C18-CrtY<br>1. pSB1C3-red <i>-as N/C, one that we use plus a digest that has worked for other groups</i><br>7. pSB3K3-red <i>-as N/C, one that we use plus a digest that has worked for other groups</i><br>42. pSB4S15-red<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>77. pSB4C15-red<br><i>-*We increased the incubation time from 30 min. to 90 min. We also added T4 DNA ligase to the already ligated plasmids we used for transformation yesterday and left them to incubate over night, since we suspect incomplete ligation may be the explanation for the bad results (digests have appeared ok on gel). Thus if transformations fail again, we can use the definitely ligated plasmids to do another immediately in the morning.</i><br><br><b>Transformation:</b><br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>75. pEL3A15-CrtB<br>76. pEL3K16-CrtI<br>78. pEL3C18-CrtY<br>42. pSB4S15-red<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>77 pSB4C15-red<br><br><b>O/N:</b><br>Lb10.1. plantarum 256 no plasmid<br>Lb11.1. plantarum 36E no plasmid <br>Lb12.1 reuteri DSM 20016 no plasmid<br><br><b>Restreak:</b><br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>57. pSB1C3-B0034-Idi<br>58. pSB1C3-B0034-ispA<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>67. pSB1C3-Cp44<br><br><b>Plasmid preparation:</b><br>7.pSB3K3<br>22.pSB1C3-B0032-BFP<br>42. pSB4S15-red<br><br><b>Gelelectrophoresis:</b><br>19. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>20. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>21. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>78. pEL3C18-CrtY<br>25. pSB1C3-CrtY<br>27. pEL3K16-red<br>51. pEL3C18-red<br>Lb13.2.3 Lc. lactis MG1363-pJP059*<br>Lb13.3 Lc. lactis MG1363-pJP059*<br>Lb13.4 Lc. lactis MG1363-pJP059*<br><i>*Of first PCR. Two PCR tubes of 13.2.3 were prepared along with one of 13.3 and one of 13.4.</i><br><br><b>Results</b><br><b>Gelelectrophoresis:</b><br><b>successful:</b><br>25. pSB1C3-CrtY<br>27. pEL3K16-red<br>51. pEL3C18-red<br><i>-Digestion Worked</i><br><br><b>failed:</b><br>19. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>20. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>21. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>78. pEL3C18-CrtY<br>Lb13.2.3 Lc. lactis MG1363-pJP059(plasmid)*<br>Lb13.3 Lc. lactis MG1363-pJP059 (colony)*<br>Lb13.4 Lc. lactis MG1363-pJP059 (colony)*<br><i>-*The gel showed nothing but primer dimer.</i><br><br><b>Digest:</b><br><b>successful:</b><br>25. pSB1C3-CrtY<br>27. pEL3K16-red<br>51. pEL3C18-red<br><br><b>Yesterdays transformation:</b><br><b>successful:</b><br>61. pSB1C3-CP1: Growth<br>61. pSB1C3-CP1 10x: Some growth<br>62. pSB1C3-CP8: Some growth<br>62. pSB1C3-CP8 10x: No growth<br>63. pSB1C3-CP11: Some growth<br>63. pSB1C3-CP11 10x: No growth<br>64. pSB1C3-CP29: Some growth<br>64. pSB1C3-CP29 10x: No growth<br>65. pSB1C3-CP30: Some growth<br>65. pSB1C3-CP30 10x: No growth<br>66. pSB1C3-CP41: Growth<br>66. pSB1C3-CP41 10x: Some growth<br>67. pSB1C3-Cp44: No growth<br>67. pSB1C3-Cp44 10x: No growth<br>68. pSB3K3-CP1-B0032-BFP: No growth<br>68. pSB3K3-CP1-B0032-BFP 10x: Growth<br>69. pSB3K3-CP8-B0032-BFP: Growth, possibly contaminated<br>69. pSB3K3-CP8-B0032-BFP 10x: No growth<br>70. pSB3K3-CP11-B0032-BFP: Growth<br>70. pSB3K3-CP11-B0032-BFP 10x: Growth<br>71. pSB3K3-CP29-B0032-BFP: Growth, olly red colonies<br>71. pSB3K3-CP29-B0032-BFP 10x: -----<br>72. pSB3K3-CP30-B0032-BFP: Growth<br>72. pSB3K3-CP30-B0032-BFP 10x: No growth<br>73. pSB3K3-CP41-B0032-BFP: Some growth, only red colonies<br>73. pSB3K3-CP41-B0032-BFP 10x: No growth<br>77.1 pSB4C15, clone 1: No growth<br>77.1 pSB4C15, clone 1 10x: No growth<br>77.2 pSB4C15, clone 2: Some growth<br>77.2 pSB4C15, clone 2 10x: No growth<br>23. pSB1C3-CrtB<br>24. pSB1C3-CrtI<br>27. pEL3K16-red<br>38. pEL3A15-red<br>51. pEL3C18-red<br><br><b>failed:</b><br>81. pSB4S15-CP41: No growth<br>82. pSB4S15-CP29: No growth<br>83. pSB4S15-CP30: No growth<br><i>→ Redo ligation and transformation.</i><br><br><b>Plasmid preparation:</b><br>7.3 pSB3K3 clone 3: 50.1 ng/µl<br>7. 4 pSB3K3 clone 4: 43.4 ng/µl<br>7. 5 pSB3K3 clone 5: 48.1 ng/µl<br>22.1.1 pSB1C3-B0032-BFP clone 1.1: 196.2 ng/µl<br>22.2.1 pSB1C3-B0032-BFP clone 2.1: 174.3 ng/µ<br>22.2.2 pSB1C3-B0032-BFP clone 2.2: 189.9 ng/µ<br>42.1 pSB4S15-red clone 1: 43.6 ng/µl<br>42.2 pSB4S15-red clone 2: 46.1 ng/µl<br><br><h2>Other experiments</h2><br>Made agarplates with chloramphenicol resistance<br>MRS-broth preparation<br>MRS-broth-agar preparation <br>MRS-broth-agar plates preparation </div>'; | ||
| + | } | ||
| + | else if(id == 'd201372') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-07-02</h1><br><b>Name of participants:</b> Christoffer Ahlström (CA), Nafisa Bashir (NB), Viktor Törnblom (VT), Anton Berglund (AB), Mikael Strandgren (MS), Viktor Blomkvist (VB), Thorsteinn O, Magnus B, Nils A, Alexander W, Christoffer F, Lovisa P, Kristoffer L, Emil M, Marcus H, Karl H, Peter C, Ken B.-A.<br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri:</b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>12. DSM 20016-noplasmid<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-noplasmid<br>11. 36E-noplasmid<br><br><b>E. faecalis:</b><br>9. JH2-2 pAMβ1<br><br><b>L. lactis:</b><br>13. MG1363-pJP059<br>15. unknown-pGus<br>16. MG1363-noplasmid<br><br><b>E.coli (strain D5α):</b> <br>42. pSB4S15-red<br>60. PK401 dam<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69.pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-Cp44-B0032-BFP<br>77. pSB4C15<br>81. pSB4S15-CP41<br>82. pSB4S15-CP29<br>83. pSB4S15-CP30<br>57. pSB1C3-B0034-Idi<br>58. pSB1C3-B0034-ispA<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br><br><br><b>Lb. delbrueckii bulgaricus:</b><br>18. Lb. delbrueckii bulgaricus<br><br><b>L. thermophilus:</b><br>19. Lc.thermophilus<br><br><h2>Todays work</h2><br><b>O/N:</b><br>Lb9.7. faecalis JH2-2 pAMβ1 clone 7 (erythromycin) <br>Lb9.8. faecalis JH2-2 pAMβ1 clone 8 (no-antibiotic)<br>Lb9.9. faecalis JH2-2 pAMβ1 clone 9 (erythromycin)<br>Lb9.10. faecalis JH2-2 pAMβ1 clone 10 (no-antibiotic) <br>Lb10.1. plantarum 256 no plasmid<br>Lb11.1. plantarum 36E no plasmid<br>Lb12.1 reuteri DSM 20016<br>Lb19.5 L. thermophilus<br>Lb19.6 L. thermophilus<br>7.3. pSB3K3-red<br>7.4. pSB3K3-red<br>7.5. pSB3K3-red<br>22.1.1. pSB1C3<br>22.2.1. pSB1C3<br>22.2.2. pSB1C3<br>42.1.1. pSB4S15-red<br>42.1.2. pSB4S15-red<br>42.1.3. pSB4S15-red<br><br><b>Digest: </b><br>Lb9.4 faecalis JH2-2 pAMβ1<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br><br><b>Gel electrophoresis: </b><br>Lb9.4 faecalis JH2-2 pAMβ1<br><br><b>Frozen stock: </b><br>Lb13.3. Lc. lactis MG1363 pJP059 clone 3<br>Lb13.4. Lc. lactis MG1363 pJP059 clone 4<br>Lb15.3. Lc. lactis MG1363 pGus clone 3<br>Lb15.4. Lc. lactis MG1363 pGus clone 4<br>Lb16.3. Lc. lactis MG1363 noplasmid clone 3<br>Lb16.4. Lc. lactis MG1363 noplasmid clone 4<br><br><b>Plasmid preparation: </b><br>Lb13.3. Lc. lactis MG1363 pJP059 clone 3<br>Lb13.4. Lc. lactis MG1363 pJP059 clone 4<br>Lb15.3. Lc. lactis MG1363 pGus clone 3<br>Lb15.4. Lc. lactis MG1363 pGus clone 4<br><br><b>PCR: </b><br>Lb13.1 Lc. lactis MG1363 pJP059<br>Lb13.2 Lc. lactis MG1363 pJP059<br>Lb13.3 Lc. lactis MG1363 pJP059<br>Lb13.4 Lc. lactis MG1363 pJP059<br>The primers for usp45 were used in order to amplify the usp45 signal peptide from the L. lactis chromosome. A gel was run to confirm fragment size.<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br><br><b>Gel electrophoresis: </b><br>Lb13.1 Lc. lactis MG1363 pJP059<br>Lb13.2 Lc. lactis MG1363 pJP059<br>Lb13.3 Lc. lactis MG1363 pJP059<br>Lb13.4 Lc. lactis MG1363 pJP059<br>1. pSB1C3-red (cut with EcoRI and SpeI today)<br>1. pSB1C3-red (cut with EcoRI and SpeI saturday)<br>7. pSB3K3-red (cut with EcoRI and PstI today)<br>22. pSB1C3-B0032-BFP (cut with XbaI and PstI saturday)<br>42. pSB4S15-red (EcoRI and SpeI today)<br>42. pSB4S15-red (cut with SalI and SacI saturday)<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br><br><b>Re-streak from frozen stock: </b><br>Lb12.1 reuteri DSM 20016 no plasmid<br>Lb12.2 reuteri DSM 20016 no plasmid<br>Two re-streaks of each clone were made on two antibiotics free plates.<br><br><b>Re-streak: </b><br>Lb19.1 L. thermophilus<br>Lb19.2 L. thermophilus<br>Lb19.3 L. thermophilus<br>Lb19.4 L. thermophilus<br>Lb19.7 L. thermophilus<br>Lb19.8 L. thermophilus<br>Lb19.9 L. thermophilus<br>Lb19.10 L. thermophilus<br><br><b>Streaking of yogurt culture: </b><br>Lb18. Lb. delbrueckii bulgaricus <br>Lb19. Lactococcus Thermophilus<br><br><b>Assembly and subcloning: </b><br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69.pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-Cp44-B0032-BFP<br>77.1 pSB4C15, clone 1<br>77.2 pSB4C15, clone 2 had less of plasmid 42 because it ran out.<br>81. pSB4S15-CP41<br>82. pSB4S15-CP29<br>83. pSB4S15-CP30<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br><br><b>Transformation: </b><br>61. pSB1C3-CP1 *<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11 *<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69.pSB3K3-CP8-B0032-BFP *<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP *<br>72. pSB3K3-CP30-B0032-BFP *<br>73. pSB3K3-CP41-B0032-BFP *<br>74. pSB3K3-Cp44-B0032-BFP<br>77. pSB4C15, clone 1<br>77.2 pSB4C15, clone 2 *<br>81. pSB4S15-CP41 *<br>82. pSB4S15-CP29<br>83. pSB4S15-CP30<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br><i>* Those marked were incubated for maybe 25 minutes longer.</i><br><br><b>Subcloning</b> of 57 and 58 into pSB1C3 (from 1. pSB1C3-Red).<br>PCR of Crt Z(with 7% DMSO), 19, 20, 21(now with VF2, VR and culture)<br><b>Gelelectroforesis</b> of CrtE, CrtZ, 19, 20, 21<br><b>Transformation</b> of 23, 24, 27, 38, 51.<br><br><h2>Results</h2><br><b>Previous day:</b><br><b>O/N: </b><br>Lb13.3 Lc. lactis MG1363 pJP059 clone 3: Good growth, NC ok!<br>Lb13.4 Lc. lactis MG1363 pJP059 clone 4: Good growth, NC ok!<br>Lb15.3 Lc. lactis MG1363 pGus clone 3: Good growth, NC ok!<br>Lb15.4 Lc. lactis MG1363 pGus clone 4: Good growth, NC ok!<br><i>→ Continue with frozen stock and plasmid preparation.</i><br><br><b>Assembly and transformation: </b><br>61. pSB1C3-CP1: Nothing<br>61. pSB1C3-CP1 10x: Nothing<br>62. pSB1C3-CP8: Growth<br>62. pSB1C3-CP8 10x: Nothing<br>63. pSB1C3-CP11: Growth<br>63. pSB1C3-CP11 10x: Nothing<br>64. pSB1C3-CP29: Growth<br>64. pSB1C3-CP29 10x: Nothing<br>65. pSB1C3-CP30: Growth<br>65. pSB1C3-CP30 10x: Nothing<br>66. pSB1C3-CP41: Nothing<br>66. pSB1C3-CP41 10x: Nothing<br>67. pSB1C3-Cp44: Growth<br>67. pSB1C3-Cp44 10x: Nothing<br>77. pSB4C15-red: Nothing <br>77. pSB4C15-red 10x: Nothing<br>45. pSB1C3-B0034-4CL Growth<br>46. pSB1C3-B0034-His-4CL Growth<br>47. pSB1C3-B0034-STS Growth<br>48. pSB1C3-B0034-His-STS Growth<br><i>→ Redo with extra care, look over the protocols, use new ligase and ligation buffer.</i><br><br><b>Today:</b><br><b>Gel electrophoresis: </b><br>43. pSB1C3-B0034-TAL Good bands<br>44. pSB1C3-B0034-His-TAL Good bands<br>Lb13.1 Lc. lactis MG1363-pJP059 <br>13.2 Lc. lactis MG1363-pJP059 <br>13.3 Lc. lactis MG1363-pJP059<br>13.4 Lc. lactis MG1363-pJP059<br>Lb9.4 faecalis JH2-2 pAMβ1 Faint blurry bands*<br>1. pSB1C3-red (cut with EcoRI and SpeI today) Good bands<br>1. pSB1C3-red (cut with EcoRI and SpeI saturday) Good bands<br>7. pSB3K3-red (cut with EcoRI and PstI today) Good bands<br>22. pSB1C3-B0032-BFP (cut with XbaI and PstI saturday) Nothing**<br>42. pSB4S15-red (EcoRI and SpeI today) Nothing**<br>42. pSB4S15-red (cut with SalI and SacI saturday) Nothing**<br><i>→ 1 and 7 were ok, they could be reused.</i><br><br><i>*The results were consistent with yesterdays Gel electrophoresis: Very faint smudged bands that lets us think there is only chromosomal DNA in this plasmid preparation.<br>**The results indicate that something may be wrong with the digests of 42. and 22., this may be one of the explanations of why we got no colonies of 77. but many of 61-67.</i><br><br><b>Plasmid preparation: </b><br>Lb13.3. Lc. lactis MG1363 pJP059 clone 3: 13.4 ng/µl<br>Lb13.4. Lc. lactis MG1363 pJP059 clone 4: 10.3 ng/µl<br>Lb15.3. Lc. lactis MG1363 pGus clone 3: 11.9 ng/µl<br>Lb15.4. Lc. lactis MG1363 pGus clone 4: 12.0 ng/µl<br><i>→ Dispose, to low concentration. Re-do a plasmid preparation from O/N with optimized conditions. </i><br><br>Gel showed fail of PCR for 19,20, 21<br>Gel showed succes of CrtE, but fail of CrtZ(no primer dimers tho)<br>Plates from yesterday showed fail of subclone/transformation<br><br><h2>Other experiments</h2><br>Made agarplates with chloramphenicol resistance<br><b>Microscopy:</b><br>Confirming Lc.thermophilus colonies on the yoghurt-plate from the previous day.<br>Confirming contamination on some plates with competent cells.<br><br>Primer design of lambda red, Crt Y, I and B both F and R<br></div>'; | ||
| + | } | ||
| + | else if(id == 'd201371') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-07-01</h1><br><b>Name of participants:</b> Lovisa P, Kristoffer L, Emil M, Marcus H, Karl H, Peter C, Christoffer, Nils, Magnus, Alexander, Anders Edlund (AE), Mikael Strandgren (MS), Christoffer Ahlström (CA), Viktor Blomkvist (VB), Viktor Törnblom(VT), Nafisa Bashir (NB), Anton Berglund (AB)<br><br><h2>Ongoing constructs:</h2><br><b>E.coli (strain D5α):</b><br>7. pSB3K3-red<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>77. pSB4C15-red<br>1. pSB1C3-red<br>60.PK401 dam-<br><br><b>L. plantarum:</b><br>Lb5. 256-rifR-pAMβ1<br><br><b>E. faecalis:</b><br>Lb9. JH2-2 pAMβ1<br><br><b>L. reuteri:</b><br>Lb12. DSM 20016-noplasmid<br><br><b>L. lactis:</b><br>Lb15 unknown-pGus<br><br><b>Lb. delbrueckii bulgaricus:</b><br>18. Lb. delbrueckii bulgaricus<br><br><b>L. thermophilus:</b><br>19. L.thermophilus<br><br><h2>Todays work</h2><br><b>Gel electrophoresis:</b><br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>Lb9.4 faecalis JH2-2 pAMβ1<br>CBP amplicon from reuteri DSM 20016<br>usp45 amplicon from lactis strain MG1363<br><br><b>PCR:</b><br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS <br>6 colonies of DSM 20016 strain reuteri<br>4 colonies of MG1363 strain lactis<br>The PCR was carried out with the CBP primers for reuteri and the usp45 primers for lactis. This in order to isolate the signal peptides from each species.<br><br><b>O/N: </b><br>Lb13. Lc. lactis MG1363 pJP059 clone 3<br>Lb13. Lc. lactis MG1363 pJP059 clone 4<br>Lb15. Lc. lactis MG1363 pGus clone 3<br>Lb15. Lc. lactis MG1363 pGus clone 4<br>7. pSB3K3-red<br><br><b>Frozen stock:</b><br>Lb5.2 plantarum 256 pAMβ1<br>Lb9. faecalis JH2-2 pAMβ1 clone 4<br>1. pSB1C3-red<br>60.PK401 dam<br><br><b>Re-streak: </b><br>Lb9. faecalis JH2-2 pAMβ1<br>Lb12. reuteri DSM20016 no plasmid<br><br><b>Plasmid preparation:</b><br>Lb5.2 plantarum 256 pAMβ1<br>Lb9. faecalis JH2-2 pAMβ1 clone 4<br>1. pSB1C3-red<br><br><b>Digest: </b><br>Lb9.4 faecalis JH2-2 pAMβ1 (To rule out chromosomal contamination in prep)<br><br><b>Assembly: </b><br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>77. pSB4C15-red<br><br><b>Transformation: </b><br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>77. pSB4C15-red*<br>*We made a transformation with one sample of 77-digest from today, and one from yesterday to try to see if the problem yesterday laid with the assembly or with the transformation<br><br><b>Streaking of yogurt culture: </b><br>18. Lb. delbrueckii bulgaricus <br>19. Lactococcus Thermophilus<br><br><b>PCR</b> of Crt E, Crt Z, 19, 20, 21, Idi<br><b>Gelelectroforisis</b> of Crt E, Crt Z, Idi <br><b>Transformation</b> of 75, 76, 78, 58<br><b>Subcloning</b> of IspA ->58<br><b>Digest</b> of Idi<br><b>Ligation</b> of 76, 78<br><br><h2>Results</h2><br><b>Previous day:</b><br><b>Assembly and transformation: </b><br>61. pSB1C3-CP1: No growth<br>62. pSB1C3-CP8: No growth<br>63. pSB1C3-CP11: No growth<br>64. pSB1C3-CP29: No growth<br>65. pSB1C3-CP30: No growth <br>66. pSB1C3-CP41: No growth<br>67. pSB1C3-Cp44: No growth<br>77. pSB4C15-red: No growth<br><i>→ Re-do all steps, but do one transformation with ligations from yesterday</i><br><br><b>Today:</b><br><b>Plasmid preparation: </b><br>Lb5.2 plantarum 256 pAMβ1 clone 2: 50.7 ng/µl<br>Lb9.4 faecalis JH2-2 pAMβ1 clone 4: 403.7 ng/µl<br>1.1 pSB1C3-red clone 1: 144.0 ng/µl<br>1.2 pSB1C3-red clone 2: 96.8 ng/µl<br><br><b>Gel electrophoresis: </b><br>Lb9.4 faecalis JH2-2 pAMβ1*<br><i>*Very faint smudges that could be a shadow from the dye. Will follow up tomorrow with another gel electrophoresis of a new digest.</i><br>CBP amplicon from reuteri DSM 20016**<br>usp45 amplicon from lactis strain MG1363**<br><i>**None of the PCRs showed the desired bands on the gel.</i><br><br><b>PCR with good results:</b><br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL (Note: weak band)<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS <br><i>- Very good results on the pcr</i><br><br>Gel of Crt E and Crt Z was failed at Ethylium Bromide bath. Redo gel<br>Gel of Idi showed succes, proceed to subclone. <br>Subclone of IspA showed only red colonies. IspA was incorrectly digested with E instead of X.<br><i>-dunno what this is! :S</i><br><br><h2>Other experiments</h2><br>M17-broth agar preparation<br>M17-broth medium preparation<br>SET-buffer preparation<br>Preparation of M17 plates</div>'; | ||
| + | } | ||
| + | else if(id == 'd2013721') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Sunday 2013-07-21</h1><p><strong>Name of participants:</strong>Hampus.E, Theodor L, Karl H, Ken B, Kristoffer L, Viktor @, Marcus H. Nils, Christoffer, Anders Edlund (AE)<br><br>'+ | ||
| + | '<h2>Ongoing constructs:</h2>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>121.pSB1C3-B0034-STS<br>121.pSB1C3-B0034-His-STS<br>'+ | ||
| + | '99. pSB1C3-CrtO<br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>123. pSB1C3-ZCD<br>125. pSB1C3-ZCD-His<br>126. pSB1C3-UGTCs2<br>127. pSB1A3-B0034-UGTCs2-His<br>129. pSB1C3-Idi (mut)<br>21. CrtE~I /w RBS, Zinc finger (PCR prod)<br>128. pSIM5-tet(Nissle)<br>130. pSB1C3-CrtE~I /w RBS, Zinc Finger<br>131. pSB4A15-J23101-CrtE~I /w RBS, Zinc Finger<br>132. pSB4A15-J23106-CrtE~I /w RBS, Zinc Finger<br>133. pSB4A15 -J23110-CrtE~I /w RBS, Zinc Finger<br>7.pSB3k3-red<br>22. B0032-BFP<br>42. pSB4S15-red<br>60. PK401 dam<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>90. BBa_J61002-J23101<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>100. pJP059<br>111. pSBLb4C15<br>112. pSBLbEc4C15<br>136.1 pSB1C3-usp45<br>'+ | ||
| + | '<strong>L. reuteri:</strong><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>14. DSM 20016-pLUL631(B?)<br><br><strong>L. plantarum:</strong><br>5. 256-rifR-pAMβ1<br><br><b>Lb. delbrueckii bulgaricus:</b><br>18. Lb. delbrueckii bulgaricus<br><br><b>Lc. thermophilus:</b><br>19. Lc.thermophilus<br><br>'+ | ||
| + | '<h2>Todays work:</h2><b>Plasmid Prep on:</b><br>117.2 pSB3T5-CP1-B0034-His-TAL<br>119.2 pSB1C3-B0034-4CL<br>120.1 pSB1C3-B0034-HIS-4CL<br>104.2 and 104.5 pSB1C3-B0034-4CL-B0034-STS<br>106.5 and 106.6 pSB3T5-J23110-B0034-TAL<br>122.2 pSB1C3-B0034-His-STS;<br><br><b>Re-streak:</b><br>131. pSB4A15-BBa-J61002-J23106-CrtE~I /w RBS, Zinc finger<br>132. pSB4A15-BBa-J61002-J23110-CrtE~I /w RBS, Zinc finger<br>133. pSB4A15-BBa-J61002-J23101-CrtE~I /w RBS, Zinc finger<br>134.1 pSB1C3-CBP<br>134.2 pSB1C3-CBP<br>134.3 pSB1C3-CBP<br>134.4 pSB1C3-CBP<br>136.1 pSB1C3-usp45<br>136.2 pSB1C3-usp45<br>136.3 pSB1C3-usp45<br>136.4 pSB1C3-usp45<br><br><b>Gel:</b><br>21. CrtE~I/w RBS,ZF<br><br><b>Overnight & screening:</b><br>99. pSB1C3-CrtO<br>128.pSIM5-tet (E.Coli Nissle) Endast O/N<br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br><br>'+ | ||
| + | '<b>Overnight:</b><br>84.1 pSB4S15-CP6<br>84.2 pSB4S15-CP6<br>86.1 pSB4S15-CP6-B0032-BFP<br>86.2 pSB4S15-CP6-B0032-BFP<br>87(1.1) pSB4S15-CP25-B0032-BFP<br>87(1.2) pSB4S15-CP25-B0032-BFP<br>88.1 pSB4S15-CP1-B0032-BFP<br>88.2 pSB4S15-CP1-B0032-BFP<br>89.1 pSB4S15-CP44-B0032-BFP<br>89.2 pSB4S15-CP44-B0032-BFP<br>97.1 pSB4S15-CP8-B0032-BFP<br>97.2 pSB4S15-CP8-B0032-BFP<br>98.1 pSB4S15-CP11-B0032-BFP<br>98.2 pSB4S15-CP11-B0032-BFP<br><br>'+ | ||
| + | '<h2>Results:</h2><br><b>Re-streak:</b><br>131. pSB4A15-BBa-J61002-J23106-CrtE~I /w RBS, Zinc finger - Unable, no clones<br>132. pSB4A15-BBa-J61002-J23110-CrtE~I /w RBS, Zinc finger - 3 clones restreaked<br>133. pSB4A15-BBa-J61002-J23101-CrtE~I /w RBS, Zinc finger - 3 clones restreaked<br><br><b>Gel</b><br>21. CrtE~I/w RBS,ZF - fail<br><br><b>Overnight & screening:</b><br>99. pSB1C3-CrtO<br>108. pSB4A15-J23106-B0034-IspA - Fail of all clones<br>109. pSB4A15-K206000-B0034-IspA - Fail of all clones<br>110. pSB4A15-K206001-B0034-IspA - Fail of all clones<br><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013720') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-07-20</h1><b>Name of participants:</b> Hampus.E, Kristoffer.L, Malin .B, Magnus, Nils, Pontus, Anders Edlund (AE), Stephanie Herman (SH)<br><br><h2>Ongoing constructs:</h2>99. pSB1C3-CrtO<br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>123. pSB1C3-ZCD<br>125. pSB1C3-ZCD-His<br>126. pSB1C3-UGTCs2<br>127. pSB1A3-B0034-UGTCs2-His<br>129. pSB1C3-Idi (mut)<br>21. CrtE~I /w RBS, Zinc finger (PCR prod)<br>128. pSIM5-tet(Nissle)<br>130. pSB1C3-CrtE~I /w RBS, Zinc Finger<br>131. pSB4A15-J23101-CrtE~I /w RBS, Zinc Finger<br>132. pSB4A15-J23106-CrtE~I /w RBS, Zinc Finger<br>133. pSB4A15 -J23110-CrtE~I /w RBS, Zinc Finger<br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>121.pSB1C3-B0034-STS<br>121.pSB1C3-B0034-His-STS<br>7.pSB3k3-red<br>22. B0032-BFP<br>42. pSB4S15-red<br>60. PK401 dam<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>90. BBa_J61002-J23101<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>100. pJP059<br>111. pSBLb4C15<br>112. pSBLbEc4C15<br>136.1 pSB1C3-usp45<br><br><b>L. reuteri:</b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br><br><b>Lb. delbrueckii bulgaricus:</b><br>18. Lb. delbrueckii bulgaricus<br><br><b>Lc. thermophilus:</b><br>19. Lc.thermophilus<br><br><h2>Todays work</h2><b>Re-streak:</b><br>84. pSB4S15-CP6<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>99. pSB1C3-CrtO<br><br><b>Overnigth:</b><br>136.1 pSB1C3-usp45<br>136.2 pSB1C3-usp45<br>111.6 pSBLb4C15-red<br>111.7 pSBLb4C15-red<br><br><b>Transformation: </b><br>131. pSB4A15-BBa-J61002-J23106-CrtE~I /w RBS, Zinc finger <br>132. pSB4A15-BBa-J61002-J23110-CrtE~I /w RBS, Zinc finger <br>133. pSB4A15-BBa-J61002-J23101-CrtE~I /w RBS, Zinc finger<br>134. pSB1C3-CBP<br>135. pSB1C3-CSP<br>136. pSB1C3-USP45<br><br><b>Plasmidprep:</b><br>109. pSB4A15-K206000-B0034-IspA<br>123. pSB1C3-ZCD<br>125. pSB1C3-ZCD-His<br>126. pSB1C3-UGTCs2<br>127. pSB1A3-B0034-UGTCs2-His<br>1.1? 90.3 j23101<br>1.2?<br>2.1?90.4 j23101<br>2.2?<br><br><b>Gel:</b><br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br><br><b>Screening pcr of constructs:</b><br>119.pSB1C3-B0034-4CL<br>120 pSB1C3-B0034-HIS-4CL<br>106 pSB3T5-J23110-B0034-TAL<br>117 pSB3T5-CP1-B0034-His-TAL<br>107 pSB3T5-CP1-B0034-TAL<br><br><b>PCR:</b><br>21. pSB4C5-pBAD/AraC-CrtE~I/w RBS, ZF (to remove promotor)<br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br><br><h2>Results</h2><b>Overnight & screening:</b><br>99. pSB1C3-CrtO<br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br><br><b>Screening of: </b><br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br><i>showed negative results on all clones but 109.2. A new screening PCR of previously un-screened clones was prepared to run overnight.</i><b>Gel<br><br>Gel succes on: </b><br>106.5 pSB3T5-J23110-B0034-TAL<br>106.6 pSB3T5-J23110-B0034-TAL<br>117.1 pSB3T5-CP1-B0034-His-TAL<br>117.2 pSB3T5-CP1-B0034-His-TAL<br>119.2 pSB1C3-B0034-4CL<br><b>No band on:</b><br>107 pSB3T5-CP1-B0034-TAL<br>116 pSB3T5-J23110-B0034-His-TAL<br>120 pSB1C3-B0034-HIS-4CL<br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013719') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-07-19</h1><br>Name of participants: Ken B-A, Kristoffer L, Malin, Magnus, Sabri, Nils, Pontus, <br>Christoffer Ahlström (CA), Stephanie Herman (SH) <br><br><h2>Ongoing constructs:</h2><b>E.coli (strain D5α): </b><br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>123. pSB1C3-ZCD<br>125. pSB1C3-ZCD-His<br>126. pSB1C3-UGTCs2<br>127. pSB1A3-B0034-UGTCs2-His<br>129. pSB1C3-Idi (mut)<br>21. CrtE~I /w RBS, Zinc finger (PCR prod)<br>128. pSIM5-tet(Nissle)<br>130. pSB1C3-CrtE~I /w RBS, Zinc Finger<br>131. pSB4A15-J23101-CrtE~I /w RBS, Zinc Finger<br>132. pSB4A15-J23106-CrtE~I /w RBS, Zinc Finger<br>133. pSB4A15 -J23110-CrtE~I /w RBS, Zinc Finger<br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br> 107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>7.pSB3k3-red<br>42. pSB4S15-red<br>60. PK401 dam<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>90. BBa_J61002-J23101<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>100. pJP059<br>111. pSBLb4C15<br>112. pSBLbEc4C15<br>136.1 pSB1C3-usp45<br><br><b>L. reuteri:</b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br><h2>Todays work</h2><b>Screening pcr of constructs:</b><br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>117. pSB3T5-CP1-B0034-His-Tal<br>119. pSB1C3-B0034-4CL (from 15)<br>120. pSB1C3-B0034-His-4CL (from 15)<br><br><b>Overnight & screening:</b><br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>123. pSB1C3-ZCD<br>125. pSB1C3-ZCD-His<br>126. pSB1C3-UGTCs2<br>127. pSB1A3-B0034-UGTCs2-His<br><br><b>Plasmid preparation:</b><br>122.2. pSB1C3-B0034-His-STS (from 15)<br>90.3. BBa_J61002 (amp)-J23101<br>90.4. BBa_J61002 (amp)-J23101<br>104.2. pSB1C3-B0034-4CL-B0034-STS <br>104.5. pSB1C3-B0034-4CL-B0034-STS<br>111.2. pSBLb4C15-E.coli ori replaced with ori from lactobacillus (pJP059) in pEL4C15<br>111.3. pSBLb4C15-E.coli ori replaced with ori from lactobacillus (pJP059) in pEL4C15<br>111.4. pSBLb4C15-E.coli ori replaced with ori from lactobacillus (pJP059) in pEL4C15<br>121.1. pSB1C3-B0034-STS (from 15)<br>121.2. pSB1C3-B0034-STS (from 15)<br><br><b>Cloning:</b><br>21. CrtE~I /w RBS, Zinc finger → 1. pSB1C3-red<br><br><b>Assembly:</b><br>21. CrtE~I /w RBS, Zinc finger, 91. BBa-J61002-J23106 → 11. pSB4A15-red <br>21. CrtE~I /w RBS, Zinc finger, 92. BBa-J61002-J23110 → 11. pSB4A15-red <br>21. CrtE~I /w RBS, Zinc finger, 103. BBa-J61002-J23101 → 11. pSB4A15-red <br><br><<b>Gel electrophoresis:</b><br>129. pSB1C3-Idi (mut)<br><br><b>Restreak</b><br>128. pSIM5-tet(Nissle)<br><br><h2>Results:</h2><b>Screening: </b><br> 108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>123. pSB1C3-ZCD<br>125. pSB1C3-ZCD-His<br>126. pSB1C3-UGTCs2<br>127. pSB1A3-B0034-UGTCs2-His<br><i>showed negative results on all clones but 109.2, which proceeded to overnight. A new screening PCR was prepared to run overnight.</i><br><br><i>showed positive results in all cases but 125.4 and 127.3. All other clones proceeded to overnight.</i><br><br><i>The overnight showed no contamination of negative controls, and growth in all clones but 127.1 and 127.2.</i><br><br><b>Transformation:</b><br>Only visual red colonies on CBP & CSP and possible 2 white colonies on usp45.<br>→ Re-do the transformation with the saved ligation solution from (18/07-13)<br>→ Re-streak the possible white colonies. <br><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013718') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><br><h1>Thursday 2013-07-18</h1><br><br><b>Name of participants:</b> Karl H, Ken B-A, Kristoffer L, Christoffer, Alexander, Magnus, Pontus,Sabri,StephanieHerman (SH), Alona Nyberg (AN), Christoffer Ahlström (CA), Viktor Blomkvist (VB)<br><br><br><h2>Ongoing constructs:</h2><b>E.coli (strain D5α):</b><br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>7.pSB3k3-red<br>22. B0032-BFP<br>42. pSB4S15-red<br>60. PK401 dam<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>90. BBa_J61002-J23101<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>100. pJP059<br>111. pSBLb4C15<br>112. pSBLbEc4C15<br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>123. pSB1C3-ZCD<br>125. pSB1C3-ZCD-His<br>126. pSB1C3-UGTCs2<br>127. pSB1A3-B0034-UGTCs2-His<br>129. pSB1C3-Idi (mut)<br>113. E.coli Nissle-pSIM5-tet<br><br><b>L. reuteri:</b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br><br><b>Lb. delbrueckii bulgaricus:</b><br>18. Lb. delbrueckii bulgaricus<br><br><b>Lc. thermophilus:</b><br>19. Lc.thermophilus<br><br><h2>Todays work</h2><b>Assembly</b><br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119. pSB1C3-B0034-4CL<br>120. pSB1C3-B0034-His-4CL<br><br><br>Ligation<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119. pSB1C3-B0034-4CL<br>120. pSB1C3-B0034-His-4CL<br><br><b>Gel-electrophoresis of screening PCR products</b><br>42. pSB4S15-red<br><br><b>Digestion:</b><br>42. pSB4S15-red<br><br><b>Transformation:</b><br>81. pSB4S15-CP41-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br><br><b>Overnight:</b> <br>90. BBa_J61002-J23101<br><br><b>Spreading of strains:</b> <br>81. pSB4S15-CP41-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br><br><b>Re-circulization:</b><br>129. pSB1C3-Idi (mut)<br><br><b>Plasmid Prep (Glycerol stock):</b><br>75. pEL3A15 - CrtB (75.1, 75.2, 75.4)<br><br><b>Transformation:</b><br>99. pSB1C3 - Crto <br>129. pSB1C3-Idi (mut)<br>113. E.coli Nissle-pSIM5-tet<br>111. pSBLb4C15<br>112. pSBLbEc4C15<br><br><b>Re-streak:</b><br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>123. pSB1C3-ZCD<br>125. pSB1C3-ZCD-His<br>126. pSB1C3-UGTCs2<br>127. pSB1A3-B0034-UGTCs2-His<br><br><br><b>Ligation of strains from yesterdays digest:</b><br>Collagen binding protein → cloned from strain 14.4<br>Cell surface protein → cloned from strain 3.1<br>usp45 → cloned from strain 13.1<br><br>*Ligated to 1.1 pSB1C3-red (From strain database 2013)<br><br><b>Transformation:</b>*<br>Collagen binding protein<br>Cell surface protein <br>usp45 <br><br>*Performed with D5(alpha) competent cells. <br><br><b>Screening:</b><br>111. pSBLb4C15<br><br><b>Spreading of strains:</b><br>Collagen binding protein [D5(alpha)] *<br>Cell surface protein [D5(alpha)] *<br>usp45 [D5(alpha)] *<br>111. pSBLb4C15 **<br>112. pSBLbEc4C15 **<br><br>*With a 10x and 1x dilution. <br><br>** 2 plates of each with 1x dilution.<br><br><h2>Results</h2><b>Re-streaks:</b><br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>123. pSB1C3-ZCD<br>125. pSB1C3-ZCD-His<br>126. pSB1C3-UGTCs2<br>127. pSB1A3-B0034-UGTCs2-His<br>were successful on all clones but 108.7.<br><br>Transformation of 129. pSB1C3-Idi (mut) did not grow.<br><br><b>Electrophoresis on digests:</b> <br>*The gel showed that the components (signal peptides and destination plasmid) had been cut and therefore have the digest succeeded. <br><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013717') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><br><h1>Thursday 2013-07-18</h1><br><br><b>Name of participants:</b> Karl H, Ken B-A, Kristoffer L, Christoffer, Sabri, Alexander, Nils, Thorsteinn, Magnus, Malin, Pontus, Niclas<br><br><br><h2>Ongoing constructs:</h2><b>E.coli (strain D5α):</b><br>75. pEL3A15 - CrtB<br>99. pSB1C3 - CrtO<br>79. pEL3K16 - CrtE <br>102. pSB1C3 - CrtE<br>101. pSB1C3 - DNA program123456<br>96. Cs42s-Promoter-ZCD-PsbA2-UGTCs2-PsbA2-Beta-carotene hydroxylase-B1002<br>57.Psb1C3-B0034-Idi<br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>123. pSB1C3-ZCD<br>125. pSB1C3-ZCD-His<br>126. pSB1C3-UGTCs2<br>127. pSB1A3-B0034-UGTCs2-His<br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>7.pSB3k3-red<br>22. B0032-BFP<br>42. pSB4S15-red<br>60. PK401 dam<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>100. pJP059<br>111. pSBLb4C15<br>112. pSBLbEc4C15<br><br><b>L. reuteri:</b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br><br><b>Lb. delbrueckii bulgaricus:</b><br>18. Lb. delbrueckii bulgaricus<br><br><b>Lc. thermophilus:</b><br>19. Lc.thermophilus<br><br><br><h2>Todays work</h2><b>Assembly</b><br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119. pSB1C3-B0034-4CL<br>120. pSB1C3-B0034-His-4CL<br><br><br><b>Ligation</b><br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119. pSB1C3-B0034-4CL<br>120. pSB1C3-B0034-His-4CL<br><br><b>Digest & Ligation:</b><br>ZCD + 1. pSB1C3-Red -> 123. pSB1C3-ZCD<br>ZCD-His + 1. pSB1C3-Red -> 125. pSB1C3-ZCD-His<br>UGTCs2 + 1. pSB1C3-Red -> 126. pSB1C3-UGTCs2<br>B0034-UGTCs2-His + 4. pSB1A3-Red -> 127. pSB1A3-B0034-UGTCs2-His<br><br><b>Digest</b><br>Collagen binding protein → cloned from strain 14.4<br>Cell surface protein → cloned from strain 3.1<br>usp45 → cloned from strain 13.1<br>1.1 pSB1C3-red (From strain database 2013)<br><br><b>Transformation:</b><br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>123. pSB1C3-ZCD<br>125. pSB1C3-ZCD-His<br>126. pSB1C3-UGTCs2<br>127. pSB1A3-B0034-UGTCs2-His<br><br><b>O/N & Screening:</b><br>75. pEL3A15 - CrtB<br>102. pSB1C3 - CrtE<br><br><b>Plasmid Prep(Glycerol stock)</b><br>79. pEL3K16 - CrtE <br>101. pSB1C3 - DNA program123456<br><br><b>PCR</b><br>21. pSB4C5 - pBAD/AraC-CrtE~I /w RBS, ZF (E,P)(To remove promotor)<br>57.pSB1C3-B0034-Idi mutagenesis (Phusion) to remove restriction site. <br><br><b>Gel-electrophoresis of screening PCR products:</b><br>111. pSBLb4C15<br>112. pSBLbEc4C15<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB4S15-CP6-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br><br><b>Sequencing preparation:</b><br>62. pSB1C3-CP8<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br><br><b>Plasmid preparation: </b><br>74. pSB3K3-CP44-B0032-BFP <br><br>Gel-electrophoresis of overnight PCR products:<br>Collagen binding protein → cloned from strain 14.4<br>Cell surface protein → cloned from strain 3.1<br>usp45 → cloned from strain 13.1<br><br><b>Re-streak: </b><br>Lb18. Lb. delbrueckii bulgaricus no plasmid<br><br><b>Spreading of strains: </b><br>Lb19. Lc.thermophilus no plasmid*<br><br>*With 1x, 10x & 50x dilution<br><br><br><h2>Results</h2>57.pSB1C3-B0034-Idi worked, gel run showed a band at suspected size , but also some smudge, probably primer dimers.<br>101. pSB1C3 - DNA program123456 worked and sent for sequence<br><br><b>Transformations:</b><br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>123. pSB1C3-ZCD<br>125. pSB1C3-ZCD-His<br>126. pSB1C3-UGTCs2<br>127. pSB1A3-B0034-UGTCs2-His<br>all grew red and white colonies, indicating successful ligation and transformation.<br><br><b>Gel-electrophoresis of overnight PCR products:</b><br>Collagen binding protein → cloned from strain 14.4: NC checks good and lines are equal to ~200bp.<br>Cell surface protein → cloned from strain 3.1: NC checks good and lines are equal to ~150bp.<br>usp45 → cloned from strain 13.1: NC checks good and lines are equal to ~140bp.<br><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013716') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><br><h1>Tuesday 2013-07-16</h1><br><br><b>Name of participants:</b> Karl H, Lovisa P, Theodor L, Ken B-A, Victor S, Hampus.E, Kristoffer L, Christoffer .F, Sabri. J, Alexander, Nils, Thorsteinn, Magnus, Malin, Pontus, Niclas, Viktor Blomkvist(VB), Alona Nyberg (AN), Christoffer Ahlström (CA)<br><br><h2>Ongoing constructs:</h2><br><br><b>E.coli (strain D5α):</b><br>7.pSB3k3-red<br>22. B0032-BFP<br>42. pSB4S15-red<br>60. PK401 dam<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>100. pJP059<br>111. pSBLb4C15<br>112. pSBLbEc4C15<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>75. pEL3A15 - CrtB<br>99. pSB1C3 - CrtO<br>79. pEL3K16 - CrtE <br>102. pSB1C3 - CrtE<br>101. pSB1C3 - DNA program123456<br>96. Cs42s-Promoter-ZCD-PsbA2-UGTCs2-PsbA2-Beta-carotene hydroxylase-B1002<br>91. BBa_J61002 (amp)-J23106<br>94. pSB1C3-K206000<br>95. pSB1C3-K206001<br>11. pSB4A15-blue(?)<br>58. pSB1C3-IspA<br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>21. pSB4C5 - pBAD/AraC-CrtE~I /w RBS, ZF<br>57. PSB1C3-B0034-Idi <br><br><b>L. reuteri:</b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>14. DSM 20016-pLUL631(B?)<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br><br><h2>Todays work</h2><b>Assembly</b><br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br><br><b>Ligation:</b><br>48. pSB1C3-B0034-His-STS<br><br><b>Gel on Digest:</b><br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>104. pSB1C3-B0034-4CL-B0034-STS<br><br><b>Restreak:</b><br>47. pSB1C3-B0034-STS<br>75. pEL3A15 - CrtB<br><br><b>Screening:</b> <br>111. pSBLb4C15<br>112. pSBLbEc4C15<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB4S15-CP6-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br><br><b>Plasmid preparation: </b><br>74. pSB3K3-CP44-B0032-BFP<br>27. pEL3K16-red<br><br><b>Overnight: </b><br>74. pSB3K3-CP44-B0032-BFP<br><br><i>Touchdown PCR with Phusion polymerase (signal peptides): CA<br>Collagen binding protein → cloned from strain 14.4<br>Cell surface protein → cloned from strain 3.1<br>usp45 → cloned from strain 13.1</i><br><br><b>Transformation:</b><br>99. pSB1C3 - CrtO<br><br><b>O/N & Screening</b><br>79. pEL3K16 - CrtE <br>102. pSB1C3 - CrtE<br>101. pSB1C3 - DNA program123456<br><br><b>Gelelectrophoresis</b><br>21. pSB4C5 - pBAD/AraC-CrtE~I /w RBS, ZF<br>57. PSB1C3-B0034-Idi (on pcr product from overnight pcr with new protocol) <br><br><b>PCR </b><br>57.PSB1C3-B0034-IDI (overnight Tuesday-Wednesday new protocol) <br><br><h2>Results</h2><b>Plasmid preparation: </b><br>74. pSB3K3-CP44-B0032-BFP: Failed! → Re-do!<br><br><b>Screening:</b> <br>111. pSBLb4C15: Failed<br>112. pSBLbEc4C15: Failed<br><br><h2>Other experiments</h2>Primer stock preparation: CA *<br>CSP_RF<br>CBP_RF<br>usp45_RF<br>CBP F<br>*Obtained a concentration of 5µM at a volume of 200µl.<br><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013715') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><br><h1>Monday 2013-07-15</h1><br><br><b>Name of participants:</b> Karl H, Lovisa P, Theodor L, Ken B-A, Victor S, Hampus.E, Christoffer, Sabri, Alexander, Nils, Thorsteinn, Magnus, Malin, Pontus, Niclas<br><br><h2>Ongoing constructs:</h2><b>E.coli (strain D5α):</b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>105. pSB3T5-J23101-B0034-TAL<br>104. pSB1C3-B0034-4CL-B0034-STS<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>79. pEL3K16 - CrtE <br>102. pSB1C3 - CrtE<br>75. pEL3A15 - CrtB<br>101. pSB1C3 - DNA program123456<br>76. pEL3K16-CrtI<br>78. pEL3C18-CrtY<br>99. pSB1C3 - CrtO<br>57.Psb1C3-B0034-Idi<br>91. BBa_J61002 (amp)-J23106<br>94. pSB1C3-K206000<br>95. pSB1C3-K206001<br>11. pSB4A15-blue(?)<br>58. pSB1C3-IspA<br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>7.pSB3k3-red<br>22. B0032-BFP<br>42. pSB4S15-red<br>60. PK401 dam<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>100. pJP059<br>111. pSBLb4C15<br>112. pSBLbEc4C15<br><br><b>L. reuteri:</b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>14. DSM 20016-pLUL631(B?)<br>22. Betacarotene-various plasmids<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-p256<br><br><h2>Todays work</h2><b>PCR Purification:</b><br>111. pSBLb4C15, digest<br>112. pSBLbEc4C15, digest<br><br><b>Ligation: </b><br>111. pSBLb4C15 (MluI+NheI) + Ori from pJP059 (MluI+NheI) <br>112. pSBLbEc4C15: (MluI+ClaI) + Ori from pJP059 (MluI+ClaI)<br><br><b>Transformation: </b><br>111. pSBLb4C15<br>112. pSBLbEc4C15<br>75. pEL3A15 - CrtB<br>99. pSB1c3 - CrtO<br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br><br><b>Overnight: </b><br>74. pSB3K3-CP44-B0032-BFP<br>101. pSB1C3 - DNA program123456 <br><br><b>Fluorescence(BFP) experiment with UV: </b><br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB4S15-CP6-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br><br><b>Plasmid preparation: </b><br>Lb10.1 plantarum 256 p256<br>Lb10.2 plantarum 256 p256<br>Lb10.3 plantarum 256 p256<br>Lb22.1 reuteri betacarotene (strain name?) various plasmids<br>44.5 pSB1C3-b0034-His-Tal<br>74. pSB3K3-CP44-B0032-BFP<br><br><b>PCR:</b><br>48. pSB1C3-B0034-His-STS<br><br><b>Gel electhrophoresis</b><br>48. pSB1C3-B0034-His-STS<br><br><b>Assembly:</b><br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br><br><b>Re-streak:</b><br>79. pEL3K16 - CrtE <br>102. pSB1C3 - CrtE<br>75. pEL3A15 - CrtB<br><br><br><b>PCR cloning of:</b><br>CrtE~I /w RBS, Zinc finger (PCR is done to remove promoter). Template should be the following construct, digested (E,P) 21. pBAD/AraC-CrtE~I /w RBS, Zinc finger <br><br>CrtE~Z /w RBS, Zinc finger (PCR is done to remove promoter). Template should be the following construct, digested (E,P) 20. pBAD/AraC-CrtE~I /w RBS, Zinc finger <br><br>CrtE~(O)-Z /w RBS, Zinc finger (PCR is done to remove promoter). Template should be the following construct, digested (E,P) 19. pBAD/AraC-CrtE~I /w RBS, Zinc finger <br><br>CrtZ (PCR is done to clone and biobrick CrtZ from the following template, 20. pBAD/AraC-CrtE~CrtZ /w RBS, Zinc Finger<br><br><b>Mutagenesis</b> <br>57. pSB1C3-B0034-Idi <br><br><b>Sequence preperation</b><br>78.1 pEL3C18-CrtY (VF and VR)<br>78.3 pEL3C18-CrtY (VF and VR)<br><br><b>Screening:</b><br>111. pSBLb4C15<br>112. pSBLbEc4C15<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB4S15-CP6-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>101. pSB1C3 - DNA program123456 <br><br><i>Touchdown PCR with Phusion polymerase (signal peptides): CA<br>Collagen binding protein → cloned from strain 14.4<br>Cell surface protein → cloned from strain 3.1<br>usp45 → cloned from strain 13.1</i><br><br><h2>Results</h2><b>PCR, Succes on:</b><br>48. pSB1C3-B0034-His-STS<br><br><b>Transformation </b><br>45. pSB1C3-B0034-4CL, failed<br>46. pSB1C3-B0034-His-4CL, failed<br>104. pSB1C3-B0034-4CL-B0034-STS, failed<br>106. pSB3T5-J23110-B0034-TAL, failed<br>108. pSB4A15-J23106-B0034-IspA, failed<br>109. pSB4A15-K206000-B0034-IspA, failed<br>110. pSB4A15-K206001-B0034-IspA, failed<br>111. pSBLb4C15<br>112. pSBLbEc4C15<br><br><i>-(45,46,104,106) Probably due to weak competent cells<br>-(111-112) No growth. Possible problems: Too short time for proper ligation, no purification before ligation. Another attempt will be madwith the ligation from yesterday and a new one with purified digests.<br><br>Nanodrop measurement of 57.Psb1C3-B0034-Idi was 80 ng/microliter.</i><br><br><b>Gel:</b><br>21 pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268, fail<br><br><b>Fluorescence(BFP) experiment with UV:</b><br>68. pSB3K3-CP1-B0032-BFP : Glowing (68.2 68.4)<br>69. pSB3K3-CP8-B0032-BFP : Glowing (69.3 69.4)<br>70. pSB3K3-CP11-B0032-BFP : Glowing (70.3)<br>71. pSB3K3-CP29-B0032-BFP : Glowing (71.1 71.2)<br>72. pSB3K3-CP30-B0032-BFP : Glowing (72.1 72.2)<br>73. pSB3K3-CP41-B0032-BFP : Glowing (73.4)<br>74. pSB3K3-CP44-B0032-BFP : Glowing (74.1.1 74.1.2)<br>81. pSB4S15-CP41-B0032-BFP : Not glowing<br>83. pSB4S15-CP30-B0032-BFP : Not glowing<br>86. pSB4S15-CP6-B0032-BFP : Not glowing<br>88. pSB4S15-CP1-B0032-BFP : Not glowing<br>89. pSB4S15-CP44-B0032-BFP : Not glowing<br><br><b>Plasmid preparation:</b><br>Lb10.1 plantarum 256 p256: 124.7 ng/µl<br>Lb10.2 plantarum 256 p256: 23.5 ng/µl<br>Lb10.3 plantarum 256 p256: 270.9 ng/µl <br>Lb22.1 reuteri betacarotene (strain name?) various plasmids: 51.4 ng/µl<br><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013714') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Sunday 2013-07-14</h1><b>Name of participants:</b> Anders Edlund (AE), Viktor Blomkvist (VB), Lovisa P, Christoffer F, Sabri J<br><br><h2>Ongoing constructs:</h2><b>E.coli (strain D5α): </b><br>7.pSB3k3-red<br>22. B0032-BFP<br>42. pSB4S15-red<br>60. PK401 dam<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>100. pJP059<br>111. pSBLb4C15<br>112. pSBLbEc4C15<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>91. BBa_J61002-J23106<br>92. BBa_J61002-J23110<br>93. BBa_J61002-J23116<br>103. pSB1k3-J23101<br>79. pEL3K16 - CrtE <br>102. pSB1C3 - CrtE<br>75. pEL3A15 - CrtB<br>101. pSB1C3 - DNA program123456<br>76. pEL3K16-CrtI<br>78. pEL3C18-CrtY<br><br><b>L. reuteri:</b><br>1. CF48-pLR581<br>2. 1063-pLUL631<br>3. DSM 20016-pVs2<br>4. 100-23-noplasmid<br>6. DSM 20016-pLUL63TsA8<br>7. DSM 20016-pGFP<br>8. 100-23-pGT232<br>14. DSM 20016-pLUL631(B?)<br>22. Betacarotene-various plasmids<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-p256<br><br><h2>Todays work</h2><b>Ligation: </b><br>111. pSBLb4C15 (MluI+NheI) + Ori from pJP059 (MluI+NheI) <br>112. pSBLbEc4C15: (MluI+ClaI) + Ori from pJP059 (MluI+ClaI)<br><br><b>Transformation:</b><br>111. pSBLb4C15<br>112. pSBLbEc4C15<br>79. pEL3K16 - CrtE <br>102. pSB1C3 - CrtE<br>75. pEL3A15 - CrtB<br><br><b>Plasmid prep. and glycerol stock:</b><br>103. pSB1k3-J23101<br>91. BBa_J61002-J23106<br>92. BBa_J61002-J23110<br>93. BBa_J61002-J23116<br>44.4 pSB1C3-b0034-His-Tal<br>76.(1-4)-pEL3K16-CrtI<br>78.(1-3)-pEL3C18-CrtY<br><br><b>Overnight:</b><br>44.5 pSB1C3-b0034-His-Tal<br>BBa_J61002-J23101 (amp)<br><br><b>Restreak:</b><br>101. pSB1C3 - DNA program123456 (Picked 4 clones)<br><br><b>Subcloning:</b><br>CrtE (PCR prod) should be digested with X,P (Can be found in digest box)<br>27. pEL3K16 - red should be digested with X,P → CrtE (PCR prod)<br>1. pSB1C3 - red should be digested with X,P → CrtE (PCR prod)<br>38. pEL3A15 - red → 23. pSB1C3-CrtB<br><br><b>PCR cloning:</b><br>CrtE~I /w RBS, Zinc finger (PCR is done to remove promoter). Template should be the following construct, digested (E,P) 21. pBAD/AraC-CrtE~I /w RBS, Zinc finger <br><br>CrtZ (PCR is done to clone and biobrick CrtZ from the following template, 20. pBAD/AraC-CrtE~CrtZ /w RBS, Zinc Finger<br><br>CrtO PCR is done to clone CrtO from the following template, digested (E,P) 19. pBAD/AraC-CrtE~Crt(O)-Crt(Z) /w RBS, Zinc finger<br><br><h2>Results</h2>Problems with the yesterday ligation, the experiment was redone<br>Transformation of 101. DNA program123456 was a sucess although 102. pSB1C3-CrtE failed<br>The PCR failed for CrtE~I, unspecific binding<br>The PCR failed for CrtZ, unspecific binding. <br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013713') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><br><h1>Saturday 2013-07-13</h1><b>Name of participants:</b> Sabri J, Nils, Kristoffer L, Theodor Löwe, Viktor Blomkvist (VB), Stephanie Herman (SH), Alona Nyberg (AN)<br><br><h2>Ongoing constructs:</h2><b>E.coli (strain D5α):</b><br>101. pSB1C3 - DNA Program123456<br>79. pEL3K16 - CrtE <br>102. pSB1C3 - CrtE<br>76. pEL3K16 - CrtI <br>78. pEL3C18 - CrtY<br>90 pSB1k3-J23101<br>91 BBa_J61002-J23106<br>92 BBa_J61002-J23110<br>93 BBa_J61002-J23116<br>44.4 pSB1C3-b0034-His-Tal<br>44.5 pSB1C3-b0034-His-Tal<br>111. pSBLb4C15: MluI+NheI<br>112. pSBLbEc4C15: MluI+ClaI<br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br><br><h2>Todays work</h2><b>Ligation:</b><br>1. pSB1C3 - red → DNA program123456 (PCR prod)<br>1. pSB1C3 - red → CrtE (PCR prod)<br>27. pEL3K16 - red → CrtE (PCR prod)<br><br><b>PCR and Gelelectrophoresis:</b><br>CrtO<br><br><b>Transformation:</b><br>101. pSB1C3 - DNA program123456<br>102. pSB1C3 - CrtE<br><br><b>Overnight, Screening:</b><br>76.(1-4) pEL3K16 - CrtI <br>78.(1-3) pEL3C18 - CrtY<br><br><b>PCR purification: </b><br>111. pSBLb4C15<br>112. pSBLbEc4C15<br><br><b>Digestion:</b><br>1. pSB1C3 - red → DNA program123456 (PCR prod)<br>111. pSBLb4C15: MluI+NheI<br>112. pSBLbEc4C15: MluI+ClaI<br>Lb13. Lc. lactis MG1363-pJP059, <i>Ori: Two experiments, Mlu1+NheI & MLuI+ClaI<br>* Parts of what will become the first shuttle-vector.</i><br><br><b>Plasmid preparation:</b><br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br><br><h2>Results</h2><b>Gel:</b><br>47. pSB1C3-B0034-STS PCR product with and without DMSO (7%)<br>Good results on GEL, both PCR with and without DMSO looks good.<br><br><img class="result-pic" src="https://static.igem.org/mediawiki/igem.org/4/43/Uppsala_gelPic_2013-07-13.jpg"><table class="table-pic-desc-table"><tr><td>1</td><td>2</td><td>3</td><td>4</td><td>5</td><td>6</td><td>7</td><td>8</td><td>9</td><td>10</td></tr><tr><td>N/C</td><td>76.1 (Sucess)</td><td>76.2 (Sucess)</td><td>76.3 (Sucess)</td><td>76.4 (Sucess)</td><td>Ladder</td><td>N/C</td><td>78.1 (Sucess)</td><td>78.2 (Sucess)</td><td>78.3 (Sucess)</td></tr><tr><td>11</td><td>12</td><td>13</td><td>14</td><td>15</td><td>16</td><td>17</td><td>18</td><td>19</td><td>20</td></tr><tr><td>Ladder</td><td>CrtE (digested with E,P)</td><td>CrtE~I (digested with X, P) (Failed)</td><td>CrtE~I (digested with E, P) (Failed)</td><td>N/C</td><td>CrtO (Failed)</td><td>ORI I (Geneticals)</td><td>ORI II (Geneticals)</td><td>Ladder</td></tr></table><br><i>CrtE PCR prod should have been digested with X,P therefore construct 79 and 102 needs to be redone </i><br><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013712') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><br><h1>Friday 2013-07-12</h1><b>Name of participants:</b> Kristoffer L, Emil M, Karl H, Peter C, Lovisa P, Theodor L, Ken B-A, Christoffer F, Sabri J, Niclas<br><br><h2>Ongoing constructs:</h2><b>E.coli (strain D5α):</b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>82. pSB4S15-CP29-B0032-BFP<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>87. pSB4S15-CP25-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>100. pJP059<br><br><b>Lb. delbrueckii bulgaricus:</b><br>18. Lb. delbrueckii bulgaricus<br><br><b>Lc. thermophilus:</b><br>19. L.thermophilus<br><br><b>L. plantarum:</b><br>5. 256-rifR-pAMβ1<br>10. 256-p256<br>11. 36E-noplasmid<br><br><b>E. faecalis:</b><br>9. JH2-2 pAMβ1<br><br><b>L. lactis:</b><br>13. MG1363-pJP059<br>15. unknown-pGus<br>16. MG1363-noplasmid<br><br><h2>Todays work</h2>Digestion:<br>22. B0032-BFP - E,P<br>11. psB4A15<br><br><b>Ligation:</b> <br>CrtE~I /w RBS + Zincfinger -> 11. pSB4A15-ed<br>CrtE(PCR-prod.) -> 27. pEL3K16-red <br>CrtE -> 1. pSB1C3 (to registry)<br><br><i>-all AMP plates from yesterday failed 90-93. (J23 series promotors)<br>Digest of construct 14 and 15 from plasmid prep with Pst1, followed by gel analysis.</i><br><br><b>Transformation:</b><br>CrtE~I /w RBS + Zincfinger -> 11. pSB4A15-ed<br>CrtE(PCR-prod.) -> 27. pEL3K16-red <br>82. pSB4S15-CP29-B0032-BFP<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>87. pSB4S15-CP25-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>100. pJP059<br>J23101<br><br>PCR on DNA program 123456<br><br>Plasmid prepp on 96.Cs42s clone nr 1 and 2. <br>Glycerol stock on 96.Cs42s clone 1 and 2.<br>Gel run on digest from plasmid preparation of 96.cs42s clone 1 and 2.<br><br><b>Re-streak:</b><br>76 pEL3K16 - CrtI (4 clones)<br>78. pEL3C18 - CrtY (3 clones)<br>74. pSB3K3-CP44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB4S15-CP6-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br><br><b>Gel electrophoresis:</b><br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>46. pSB1C3-B0034-His-4CL<br>48. pSB1C3-B0034-His-STS<br>pSB4C15 PCR-product with intact ori<br>pSB4C15 PCR-product without ori<br><br><b>Overnight culture:</b><br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>Lb10.3 plantarum 256 p256<br>Lb10.4 plantarum 256 p256<br><br><b>Spreading:</b><br>87. pSB4S15-CP25-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>Lb10.3 plantarum 256 p256<br>Lb10.4 plantarum 256 p256<br><br><h2>Results</h2><b>Transformation:</b><br>Successful:<br>76 pEL3K16-CrtI<br>78 pEL3C18-CrtY<br> <br>failed:<br>75 pEL3A15-CrtB<br><br>The result from nanodrop measurement of 96.Cs42s clone 1 was 408.3 nanogram/microliter and clone 2 was 332. Also both had a god a260/a280 ratio.<br>The result from the gel run of the two 96.Cs42s clones indicates an construct with a size of ~4000 bp. <br><br><b>Gel on PCRs:</b><br>Successful:<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br><br>failed<br>47. pSB1C3-B0034-STS<br>46. pSB1C3-B0034-His-4CL<br>48. pSB1C3-B0034-His-STS<br><br><i>-PCR-amplification: 47. pSB1C3-B0034-STS, change temperature -2 degrees<br><br>Construct 14 had two bands on the gel, indicating illegal restriction sites. Construct 15 only had one band and no indication of illegal restriction sites.</i><br><br><h2>Other experiments</h2>Casting LB-agar plates: AB<br>27 plates containing spectinomycin.<br><br><b>Preparation for sequencing:</b><br>69.1 pSB3K3-CP8-B0032-BFP<br>71.3 pSB3K3-CP29-B0032-BFP<br>72.3 pSB3K3-CP30-B0032-BFP<br><br><b>Dilution of primers: </b><br>200 µl VF2 (5 µM)<br>200 µl VR (5 µM)<br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013711') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><br><h1>Thursday 2013-07-11</h1><br><b>Name of participants:</b> Kristoffer L, Emil M, Karl H, Peter C, Lovisa P, Theodor L, Ken B-A, Christoffer Ahlström (CA), Anton Berglund (AB), Stephanie Herman (SH), Alona Nyberg (AN), Viktor Törnblom (VT), Viktor Blomkvist (VB), Mikael Strandgren (MS), Niclas, Sabri , Nils, Christoffer<br><br><h2>Ongoing constructs:</h2><br><b>E.coli (strain D5α): </b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>96. Cs42s <br><br><b>L. plantarum:</b><br>10. 256-p256<br><br><h2>Todays work</h2><br><b>Transformation:</b> <br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>77. pSB4C15-red*<br>75. pEL3A15 - CrtB<br>76 pEL3K16 - CrtI<br>78. pEL3C18 - CrtY<br><br><b>Restreaks:</b> <br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>Lb10.3 plantarum 256 p256 *<br>Lb10.4 plantarum 256 p256<br>Lb10.5 plantarum 256 p256 *<br>Lb10.6 plantarum 256 p256<br><br><b>PCR:</b><br>77.(1-4) pSB4C15-red*<br><i>*Two different amplifications of plasmid adding restriction-sites with different primers.<br> One product with no removals and one without ori.</i><br><br><b>PCR-amplification:</b><br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br><br><b>Digest:</b><br>14 pET13b(amp)-zifz:4cl-PBSII:STS<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br><i>-from plasmid prep with Pst1.</i><br><br><b>PCR screening & gel extraction:</b><br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br><br><b>Ligation: </b><br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br><br><b>Plasmid preparation:</b><br>77. pSB4C15-red<br><br>PCR screening of cs42s with RedTaq. Overnight culture on transfrormed 96.Cs42s.<br>Prepare of Vf2 and VR primer solution, 20x. 5 tubes each with 200 microliter/tube.<br><br><b>Subcloning of the following CrtB, CrtI, CrtY</b><br><br>23. pSB1C3-CrtB → pEL3A15 - red<br>24. pSB1C3-CrtI → pEL3K16 - red<br>25. CrtY(PCR prod.) → pEL3C18 - red<br><br><h2>Results</h2><br><b>Plasmid preparation: </b><br>77.1 pSB4C15-red: 47.2 ng/µl<br>77.2 pSB4C15-red: 50.2 ng/µl<br>77.3 pSB4C15-red: 44.0 ng/µl<br>77.4 pSB4C15-red: 54.4 ng/µl<br><br><b>Screening:</b><br>didn’t look good, the pcr product was approximatley 1000 bp on the gel.<br> Should have been around 4000.<br><br><h2>Other experiments</h2><br><b>Sequence verification:</b><br>61.1 pSB1C3-CP1<br>63.1 pSB1C3-CP11<br>67.1 pSB1C3-Cp44<br><br><b>Casting LB-agar plates: </b><br>26 plates containing spectinomycin.<br><br><b>Preparation of LB-agar solution: </b><br><b>transformation from kit:</b><br>90. BBa_J61002 (amp)-J23101<br>91. BBa_J61002 (amp)-J23106<br>92. BBa_J61002 (amp)-J23110<br>93. BBa_J61002 (amp)-J23116<br><br><b>Restreek from stock:</b><br>2012 SD(Strain Database) 13 and 61(promotors).<br>Aliquoting VF2- and VR-primers.<br><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013710') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><br><h1>Wednesday 2013-07-10</h1><br><b>Name of participants:</b> Kristoffer L, Emil M, Karl H, Peter C., Lovisa P, Ken B-A, Alexander, Niclas, Magnus, Nils, Thorsteinn, Christoffer Ahlström (CA), Anton Berglund (AB), Stephanie Herman (SH), Viktor Törnblom (VT)<br><br><h2>Ongoing constructs:</h2><br><b>E.coli (strain D5α):</b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>96. p?A?-Cs42s<br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-Cp44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB3K3-CP44-B0032-BFP<br>87. pSB3K3-CP1-B0032-BFP<br>88. pSB3K3-CP41-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br><br><h2>Todays work</h2><br><b>Digestion:</b> <br>22. pSB1C3-RBS-B0032-BFP<br><br><b>Ligation:</b><br>44. pSB1C3-B0034-His-TAL<br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-Cp44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB3K3-CP44-B0032-BFP<br>87. pSB3K3-CP1-B0032-BFP<br>88. pSB3K3-CP41-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br><br><b>transformation:</b><br>44. pSB1C3-B0034-His-TAL<br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-Cp44-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>86. pSB3K3-CP44-B0032-BFP<br>87. pSB3K3-CP1-B0032-BFP<br>88. pSB3K3-CP41-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br><br><b>Re-streak</b> <br>96. p?A?-cs42s<br><i>-Streak of transformations of ligated 20 and positive controll (AV,NA)<br>PCR of 19(digested EP), 20(digested EP), 21(digested EP) and CrtO(from 21digestEP) (AV,NA)</i><br><br><b>Spreading of strains: </b><br>Lb4. reuteri 100-23 no plasmid<br>Lb12. reuteri DSM 20016 no plasmid<br>Lb10. plantarum 256 p256<br>Lb11. plantarum 36E no plasmid<br>Lb10. plantarum 256 noplasmid: Growth<br>Lb11. plantarum 36E noplasmid: Growth<br>→ Spread out (1x) on erythromycin plates, to examine whether they are naturally resistant.<br><br><h2>Results</h2><br><b>Transformation: </b> <br>Lb4. reuteri 100-23 noplasmid: Growth <br>Lb12. reuteri DSM 20016 noplasmid: Growth<br><br><b>Re-streak:</b> <br>96. p?A?-cs42s worked well. <br><br><b>O/N culture:</b><br>69.1 pSB3K3-CP8-B0032-BFP <br>69.2 pSB3K3-CP8-B0032-BFP <br>69.3 pSB3K3-CP8-B0032-BFP <br>69.4 pSB3K3-CP8-B0032-BFP <br>70.1 pSB3K3-CP11-B0032-BFP <br>70.2 pSB3K3-CP11-B0032-BFP <br>70.3 pSB3K3-CP11-B0032-BFP <br>70.4 pSB3K3-CP11-B0032-BFP <br>71.1 pSB3K3-CP29-B0032-BFP<br>71.2 pSB3K3-CP29-B0032-BFP<br>71.3 pSB3K3-CP29-B0032-BFP<br>71.4 pSB3K3-CP29-B0032-BFP<br>72.1 pSB3K3-CP30-B0032-BFP<br>72.2 pSB3K3-CP30-B0032-BFP<br>72.3 pSB3K3-CP30-B0032-BFP<br>72.4 pSB3K3-CP30-B0032-BFP<br>*Growth<br><br><b>Plasmid preparation: </b><br>69.1 pSB3K3-CP8-B0032-BFP: 80.5 ng/µl<br>69.2 pSB3K3-CP8-B0032-BFP: 75.7 ng/µl<br>69.3 pSB3K3-CP8-B0032-BFP: 60.5 ng/µl<br>69.4 pSB3K3-CP8-B0032-BFP: 50.8 ng/µl<br>70.1 pSB3K3-CP11-B0032-BFP: 71.7 ng/µl<br>70.2 pSB3K3-CP11-B0032-BFP: 56.3 ng/µl<br>70.3 pSB3K3-CP11-B0032-BFP: 56.0 ng/µl<br>70.4 pSB3K3-CP11-B0032-BFP: 45.3 ng/µl<br>71.1 pSB3K3-CP29-B0032-BFP: 54.0 ng/µl<br>71.2 pSB3K3-CP29-B0032-BFP: 49.1 ng/µl<br>71.3 pSB3K3-CP29-B0032-BFP: 83.0 ng/µl<br>71.4 pSB3K3-CP29-B0032-BFP 67.8 ng/µl<br>72.1 pSB3K3-CP30-B0032-BFP 53.8 ng/µl<br>72.2 pSB3K3-CP30-B0032-BFP 71.8 ng/µl<br>72.3 pSB3K3-CP30-B0032-BFP 109.6 ng/µl<br>72.4 pSB3K3-CP30-B0032-BFP 106.8 ng/µl<br></div>'; | ||
| + | } | ||
| + | else if(id == 'd201379') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><br><h1>Tuesday 2013-07-09</h1><br><b>Name of participants:</b> Kristoffer L, Emil M, Marcus H, Karl H, Peter C, Ken B.-A., Lovisa P, Theodor L, Nils, Alexander, Christoffer, Niclas, Magnus, Thorsteinn, Christoffer Ahlström (CA), Stephanie Herman (SH), Nafisa Bashir (NB), Anton Berglund (AB), Mikael Strandgren (MS), Viktor Blomkvist (VB), Viktor Törnblom (VT)<br><br><h2>Ongoing constructs:</h2><br><b>E.coli (strain D5α): </b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>96. p?-Cs42s<br>81. pSB4S15-CP41<br>83. pSB4S15-CP30<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>86. pSB3K3-CP44<br>87. pSB3K3-CP1<br>88. pSB3K3-CP41<br>89. pSB4S15-CP44<br>90. pSB4S15-CP1<br>69. pSB3K3-CP8-B0032-BFP <br>70. pSB3K3-CP11-B0032-BFP <br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br><br><b>L. reuteri:</b><br>Lb4. 100-23-no plasmid<br>Lb12. DSM 20016-no plasmid<br>Lb10. 256-noplasmid<br>LB11. 36E-noplasmid<br><br><h2>Todays work</h2><br><b>Gel analysis:</b><br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br><br><b>Ligation:</b><br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br><br><b>OD600 measurement: </b><br>Lb4. reuteri 100-23 no plasmid<br>Lb12. reuteri DSM 20016 no plasmid<br>Lb10. plantarum 256 no plasmid<br>Lb11. plantarum 36E no plasmid<br><br><b>Competent cells preparation:</b><br>Lb4. reuteri 100-23 no plasmid<br>Lb12. reuteri DSM 20016 no plasmid<br>Lb10. plantarum 256 no plasmid<br>Lb11. plantarum 36E no plasmid<br><br><b>Transfromation:</b><br>Lb4. reuteri 100-23 no plasmid<br>Lb12. reuteri DSM 20016 no plasmid<br>Lb10. plantarum 256 no plasmid<br>Lb11. plantarum 36E no plasmid<br><br><b>Re-streaks:</b><br>77. pSB4C15-red<br><br><b>Transformation:</b><br>81. pSB4S15-CP41<br>83. pSB4S15-CP30<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>86. pSB3K3-CP44<br>87. pSB3K3-CP1<br>88. pSB3K3-CP41<br>89. pSB4S15-CP44<br>90. pSB4S15-CP1<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br><br><b>O/N culture:</b><br>69.1 pSB3K3-CP8-B0032-BFP <br>69.2 pSB3K3-CP8-B0032-BFP <br>69.3 pSB3K3-CP8-B0032-BFP <br>69.4 pSB3K3-CP8-B0032-BFP <br>70.1 pSB3K3-CP11-B0032-BFP <br>70.2 pSB3K3-CP11-B0032-BFP <br>70.3 pSB3K3-CP11-B0032-BFP <br>70.4 pSB3K3-CP11-B0032-BFP <br>71.1 pSB3K3-CP29-B0032-BFP<br>71.2 pSB3K3-CP29-B0032-BFP<br>71.3 pSB3K3-CP29-B0032-BFP<br>71.4 pSB3K3-CP29-B0032-BFP<br>72.1 pSB3K3-CP30-B0032-BFP<br>72.2 pSB3K3-CP30-B0032-BFP<br>72.3 pSB3K3-CP30-B0032-BFP<br>72.4 pSB3K3-CP30-B0032-BFP<br><br>Elution and transformation of 96. The transfrormed samples were plated (80μl of 1x and 200μl of 10x dilution) on all available resistances, as the backbone was unknown.<br>Ligation of 20 with all diffrent combinations of ligase buffer and ligase. (AV, CF, NA)<br>Transformation of ligation mentioned above, and a positive controll of 20 plasmidprepp. Although plates for streaking were missing so we couldn’t streak (CF, AV, NA)<br><br><h2>Results</h2><br><b>O/N:</b><br>Lb4. reuteri 100-23 no plasmid (clone 1): NC ok! → follow protocol for transformation<br>Lb12. reuteri DSM 20016 no plasmid (clone 1): NC ok! → follow protocol for transformation, watch in microscope for contamination<br>Lb10. plantarum 256 no plasmid (clone 1): NC ok! → follow protocol for transformation<br>Lb11. plantarum 36E no plasmid (clone 1): NC ok! → follow protocol for transformation<br><br><b>Transformation:</b><br>77. pSB4C15-red: Growth on Cm, some red colonies, some blue, many white (?!)<br><br><b>OD600 measurement:</b><br>Lb4. reuteri 100-23 no plasmid: 0.195 (Measure) & 0.668 (Cultivate) → proceed<br>Lb12. reuteri DSM 20016 no plasmid: 0.179 (Measure) & 0.680 (Cultivate) --”--<br>Lb10. plantarum 256 no plasmid: 0.115 (Measure) & 0.724 (Cultivate) --”--<br>Lb11. plantarum 36E no plasmid: 0.148 (Measure) & 0.624 (Cultivate) --”--<br><br><b>Microscopy: </b><br>Lb12. reuteri DSM 20016 no plasmid: No contamination was sighted! → proceed!<br><br><b>transformation:</b><br>Successful:<br>45. pSB1C3-B0034-4CL<br>96. p?-Cs42s<br><br><b>failed:</b><br>44. pSB1C3-B0034-His-TAL<br><h2>Other experiments</h2>Cold buffer preparation (1 l) <br><br>50% glycerol stock preparation (100 ml)<br><br>Microscopy: Lb12. reuteri DSM 20016 no plasmid<br></div>'; | ||
| + | } | ||
| + | else if(id == 'd201378') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-07-08</h1><br><b>Name of participants:</b> Alexander, Nils, Christofffferz , Magnus, Niclas, Thorsteinn, Kristoffer L, Emil M, Marcus H, Karl H, Peter C, Ken B.-A., Lovisa P<br><br><h2>Ongoing constructs</h2><br><b>E.coli (strain D5α): </b><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>57. pSB1C3-B0034-Idi<br>58. pSB1C3-B0044-IspA<br>81. pSB4S15-CP41<br>83. pSB4S15-CP30<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>86. pSB3K3-CP44<br>87. pSB3K3-CP1<br>88. pSB3K3-CP41<br>89. pSB4S15-CP44<br>90. pSB4S15-CP1<br><br><b>L. reuteri:</b><br>Lb4. 100-23-no plasmid<br>Lb12. DSM 20016-no plasmid<br><br><b>L. plantarum:</b><br>Lb10. 256-noplasmid<br>LB11. 36E-noplasmid<br><br><h2>Todays work</h2><br><b>PCR amplification and gel analysis:</b><br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br><br><b>Plasmid prep.:</b> <br>47. pSB1C3-B0034-STS<br>Clone: 47.9 and 47.20<br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br><br><b>Sequencing prep.:</b><br>47. pSB1C3-B0034-STS <br>Clone: 47.9 and 47.20<br>48. pSB1C3-B0034-His-STS<br>Clone: 48.1<br><br><b>Gel extraction:</b><br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br><br><b>Restreak: </b><br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br><br><b>Digest</b><br>PCR products from succefull gel extraction.<br><br><b>Ligation: </b><br>81. pSB4S15-CP41<br>83. pSB4S15-CP30<br>84. pSB4S15-CP6<br>85. pSB4S15-CP25<br>86. pSB3K3-CP44<br>87. pSB3K3-CP1<br>88. pSB3K3-CP41<br>89. pSB4S15-CP44<br>90. pSB4S15-CP1<br><br><br><b>Frozen stock: </b><br>Lb4. reuteri 100-23 no plasmid (clone 1 & 2)<br>Lb12. reuteri DSM 20016 no plasmid (clone 1 & 2)<br>Lb10. plantarum 256 noplasmid (clone 1 & 2)<br>Lb11. plantarum 36E no plasmid (clone 1 & 2)<br><br><b>O/N:</b> <br>Lb4. reuteri 100-23 no plasmid (clone 1)<br>Lb12. reuteri DSM 20016 no plasmid (clone 1)<br>Lb10. plantarum 256 no plasmid (clone 1)<br>Lb11. plantarum 36E no plasmid (clone 1)<br><br><b>Screening:</b><br>43. pSB1C3-B0034-TAL<br><br><b>O/N culture:</b><br>43.7. pSB1C3-B0034-TAL<br><br><b>Subcloning:</b><br>23->4, 24->4, 25->4(kontroll), 23->38, 24->27, 25->51 (CF, AV)<br>Transformation of 23->4, 24->4, 25->4(kontroll), 23->38, 24->27, 25->51 (CF, AV)<br>Primer design and order of lambda red (NA)<br>Sequencing preparation of 57.3 and 57.5 (pSB1C3-B0034-Idi), 58.1 and 58.4 (pSB1C3-B0034-IspA)<br><br><h2>Results</h2><br><b>Microscopy:</b> *<br><i>*No contamination was seen, save plates for later. → Proceed with O/N of two clones from the no-plasmid strains.</i> <br><br>Primers for lambda red and Cs42s were ordered today :D.<br>Transformation of 23->4(blank), 24->4(blank), 25->4(blank), 23->38(blank), 24->27(a few clones, both red and white on x1), 25->51(blank)<br><br><b>PCR amplification:</b><br>43-46 failed, most likely primer dimers - retry with different concentrations of template DNA, higher concentration of DMSO and a colony PCR of each construct.<br><br><b>gel extraction: </b><br><b>Successfull </b><br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br><br><b>failed</b><br>43. pSB1C3-B0034-STS<br>46. pSB1C3-B0034-His-4CL<br><br><b>Screening:</b><br>one succefull clone - 43.7 pSB1C3-B0034-STS<br><br><h2>Other experiments</h2><br><b>Microscopy:</b> CA*<br>*Of plates from 04/07 and O/N from 07/07 to see if contaminated. </div>'; | ||
| + | } | ||
| + | else if(id == 'd201377') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Sunday 2013-07-07</h1><br><b>Name of participants:</b>Anders Edlund (AE)<br><br><h2>Ongoing constructs</h2><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>77. pSB4S15-C-resistance<br><br><h2>Todays work</h2><br><b>Transformation (AE)</b><br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>77. pSB4S15-C-resistance<br></div>'; | ||
| + | } | ||
| + | else if(id == 'd201376') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-07-06</h1><br><b>Name of participants:</b> Stephanie Herman (SH), Anders Edlund (AE), Nafisa Bashir (NB)<br><br><h2>Ongoing constructs:</h2><br><b>E.coli (strain D5α):</b><br>42. pSB4S15-red<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>68. pSB3K3-CP1-B0032-BFP<br>69.pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-Cp44-B0032-BFP<br>77. pSB4C15<br><br><h2>Todays work</h2><br><b>Plasmid Preparation:</b> <br>42. pSB4S15-red<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br><br><b>Transformation: </b><br>68. pSB3K3-CP1-B0032-BFP<br>69.pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-Cp44-B0032-BFP<br>77. pSB4C15<br><br><h2>Results</h2><br><b>Previous day:</b><br>The O/N on the nonplasmids 4, 10, 11, 12 from the previous day had grown at a previously unexperienced rate. A contamination of the original plates from Stefan Roos was suspected as the NC was clear. The O/N was put in the cold room awaiting examination by microscope.<br><br><b>Today:</b><br><b>Plasmid Preparation:</b><br>42. pSB4S15-red<br>61. pSB1C3-CP1: (1) 203.2, (2) 202.9<br>62. pSB1C3-CP8: (1) 254.5, (2) 229.2<br>63. pSB1C3-CP11: (1) 165.0, (2) 260.1<br>64. pSB1C3-CP29: (1) 315.4, (2) 221.9<br>65. pSB1C3-CP30: (1) 243.1, (2) 101.5<br>66. pSB1C3-CP41: (1) 190.7, (2) 68.4<br>67. pSB1C3-Cp44: (1) 169.1, (2) 188.7<br></div>'; | ||
| + | } | ||
| + | else if(id == 'd201375') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-07-05</h1><br><b>Name of participants:</b> Kristoffer L, Emil M, Marcus H, Karl H, Peter C, Ken B.-A., Christoffer Ahlström (CA), Anton Berglund (AB), Stephanie Herman (SH), Mikael Strandgren (MS), Viktor Blomkvist (VB), Nafisa Bashir (NB), Magnus, Thorsteinn, Alexander, Nils, Christoffer<br><br><h2>Ongoing constructs:</h2><br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-CP44<br>57. pSB1C3-B0034-Idi<br>58. pSB1C3-B0034-ispA<br><br><b>L. reuteri</b><br>Lb4. 100-23 no plasmid<br>Lb12. DSM 20016 no plasmid<br><br><b>L. plantarum:</b><br>Lb10. 256 no plasmid<br>Lb11. 36E no plasmid<br><br><b>L. thermophilus:</b><br>Lb19. L.thermophilus<br><br><b>Todays work</b><br>Screening PCR & gel analysis of constructs:<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>61.1 pSB1C3-CP1<br>61.2 pSB1C3-CP1<br>62.1 pSB1C3-CP8<br>62.2 pSB1C3-CP8<br>62.3 pSB1C3-CP8<br>62.4 pSB1C3-CP8<br>63.1 pSB1C3-CP11<br>63.2 pSB1C3-CP11<br>63.3 pSB1C3-CP11<br>63.4 pSB1C3-CP11<br>64.1.1 pSB1C3-CP29<br>64.1.2 pSB1C3-CP29<br>64.1.3 pSB1C3-CP29<br>64.1.4 pSB1C3-CP29<br>65.1.1 pSB1C3-CP30<br>65.1.2 pSB1C3-CP30<br>65.2.1 pSB1C3-CP30<br>65.2.2 pSB1C3-CP30<br>66.5 pSB1C3-CP41<br>66.6 pSB1C3-CP41<br>66.7 pSB1C3-CP41<br>66.8 pSB1C3-CP41<br>67.1.1 pSB1C3-CP44<br>67.1.2 pSB1C3-CP44<br>67.2.1 pSB1C3-CP44<br>67.2.2 pSB1C3-CP44<br><br><b>Plasmid prep & glycerol stock:</b><br>48. pSB1C3-B0034-His-STS<br><br><b>O/N:</b> <br>57. pSB1C3-B0034-Idi<br>58. pSB1C3-B0034-ispA<br>61. pSB1C3-CP1<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>66. pSB1C3-CP41<br>67. pSB1C3-Cp44<br>47.(9, 20) pSB1C3-B0034-STS<br>Lb4. reuteri 100-23 no plasmid<br>Lb10. plantarum 256 no plasmid<br>Lb11. plantarum 36E no plasmid<br>Lb12. reuteri DSM 20016 no plasmid<br><br><b>Re-streaks:</b><br>Lb19. L.thermophilus<br><br><h2>Results</h2><br>The O/N on 42. pSB4S15-red was done improperly in 15 ml falcon tubes and the lids were screwed on too tightly. A new O/N was done on 42. pSB4S15-red by NB.<br><br><b>PCR screen and gel electrophoresis:</b><br>61.1 pSB1C3-CP1<br>61.2 pSB1C3-CP1<br>62.1 pSB1C3-CP8<br>62.2 pSB1C3-CP8<br>62.3 pSB1C3-CP8<br>62.4 pSB1C3-CP8<br>63.1 pSB1C3-CP11<br>63.2 pSB1C3-CP11<br>63.3 pSB1C3-CP11<br>63.4 pSB1C3-CP11<br>64.1.1 pSB1C3-CP29<br>64.1.2 pSB1C3-CP29<br>64.1.3 pSB1C3-CP29<br>64.1.4 pSB1C3-CP29<br>65.1.1 pSB1C3-CP30<br>65.1.2 pSB1C3-CP30<br>65.2.1 pSB1C3-CP30<br>65.2.2 pSB1C3-CP30<br>66.5 pSB1C3-CP41<br>66.6 pSB1C3-CP41<br>66.7 pSB1C3-CP41<br>66.8 pSB1C3-CP41<br>67.1.1 pSB1C3-CP44<br>67.1.2 pSB1C3-CP44<br>67.2.1 pSB1C3-CP44<br>67.2.2 pSB1C3-CP44<br><br>*The gel only showed smudgy bands.<br><br><b>succesful</b><br>47.9 pSB1C3-B0034-STS<br>47.20 pSB1C3-B0034-STS<br><br><b>failed</b><br>43. pSB1C3-B0034-TAL<br>46. pSB1C3-B0034-His-4CL<br><br>No contamination in negative control -_-<br>Religation from yesterday failed. No colonies, only a few at 76, both red and white. <br><br><h2>ther experiments</h2><br><b>Contamination control:</b> CA *<br><br><i>*Contamination control of growth medium for Lc.lactis and Lb.reuteri.<br> Two test from both bottles. </i><br><br><b>E.coli growth test:</b> CA *<br><br><i>*To see if strain 1 of e.coli in strain database 13 grow on M17 and MRS plates as well. </i></div>'; | ||
| + | } | ||
| + | else if(id == 'd201374') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-07-04</h1><b>Name of participants:</b> Lovisa P, Kristoffer L, Emil M, Marcus H, Karl H, Peter C, Ken B.-A., Christoffer Ahlström (CA), Nafisa Bashir (NB), Mikael Strandgren (MS), Anton Berglund (AB), Magnus, Thorsteinn, Alexander, Nils, Christoffer<br><br><h2>Ongoing constructs</h2><b>E.coli (strain D5α):</b> <br>15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br>57. pSB1C3-B0034-Idi<br>58. pSB1C3-B0034-ispA<br>42. pSB4S15-red<br>62. pSB1C3-CP8<br>63. pSB1C3-CP11<br>64. pSB1C3-CP29<br>65. pSB1C3-CP30<br>67. pSB1C3-CP44<br>68. pSB3K3-CP1-B0032-BFP<br>69.pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br><br><b>L. lactis:</b><br>Lb13. MG1363-pJP059<br>Lb15. unknown-pGus<br><br><b>E. faecalis:</b><br>Lb9. JH2-2 pAMβ1<br><br><h2>Todays work</h2><b>Frozen stock and plasmid preparation:</b> <br>Lb9.7. faecalis JH2-2 pAMβ1 clone 7 (erythromycin) <br>Lb9.8. faecalis JH2-2 pAMβ1 clone 8 (no-antibiotic)<br>Lb9.9. faecalis JH2-2 pAMβ1 clone 9 (erythromycin)<br>Lb9.10. faecalis JH2-2 pAMβ1 clone 10 (no-antibiotic) <br>Lb13.5. Lc. lactis MG1363 pJP059 clone 5<br>Lb13.6. Lc. lactis MG1363 pJP059 clone 6<br>Lb13.7. Lc. lactis MG1363 pJP059 clone 7<br>Lb13.8. Lc. lactis MG1363 pJP059 clone 8<br>Lb15.5. Lc. lactis MG1363 pGus clone 5<br>Lb15.6. Lc. lactis MG1363 pGus clone 6<br>Lb15.7. Lc. lactis MG1363 pGus clone 7<br>Lb15.8. Lc. lactis MG1363 pGus clone 8<br><br><b>Touchdown PCR: </b><br>Lb13.2.2 Lc. lactis MG1363 pJP059<br>Lb13.2.3 Lc. lactis MG1363 pJP059<br>Lb13.3 Lc. lactis MG1363 pJP059<br>Lb13.4 Lc. lactis MG1363 pJP059<br><br><b>Screening PCR: </b><br>62.1 pSB1C3-CP8<br>62.2 pSB1C3-CP8<br>63.1 pSB1C3-CP11<br>63.2 pSB1C3-CP11<br>63.3 pSB1C3-CP11<br>63.4 pSB1C3-CP11<br>64.1.1 pSB1C3-CP29<br>64.1.2 pSB1C3-CP29<br>64.1.3 pSB1C3-CP29<br>64.1.4 pSB1C3-CP29<br>65.1.1 pSB1C3-CP30<br>65.1.2 pSB1C3-CP30<br>65.2.1 pSB1C3-CP30<br>65.2.2 pSB1C3-CP30<br>67.1.1 pSB1C3-CP44<br>67.1.2 pSB1C3-CP44<br>67.2.1 pSB1C3-CP44<br>67.2.2 pSB1C3-CP44<br><br><b>Gel electrophoresis:</b> <br>Lb13.2.2 lactis MG1363 pJP059 (PCR with DMSO done yesterday)<br>Lb13.2.3 lactis MG1363 pJP059 (PCR with DMSO done yesterday)<br>Lb13.3 lactis MG1363 pJP059 (PCR with DMSO done yesterday)<br>Lb13.4 Lc. lactis MG1363 pJP059 (PCR with DMSO done yesterday)<br>Lb13.2.2 Lc. lactis MG1363 pJP059 (Touchdown-PCR today)<br>Lb13.2.3 Lc. lactis MG1363 pJP059 (Touchdown-PCR today)<br>Lb13.3 Lc. lactis MG1363 pJP059 (Touchdown-PCR today)<br>Lb13.4 Lc. lactis MG1363 pJP059 (Touchdown-PCR today)<br>62.1 pSB1C3-CP8 (Screening-PCR today, was run two times)<br>62.2 pSB1C3-CP8 (Screening-PCR today, was run two times)<br>63.1 pSB1C3-CP11 (Screening-PCR today, was run two times)<br>63.2 pSB1C3-CP11 (Screening-PCR today, was run two times)<br>63.3 pSB1C3-CP11 (Screening-PCR today, was run two times)<br>63.4 pSB1C3-CP11 (Screening-PCR today, was run two times)<br>64.1.1 pSB1C3-CP29 (Screening-PCR today, was run two times)<br>64.1.2 pSB1C3-CP29 (Screening-PCR today, was run two times)<br>64.1.3 pSB1C3-CP29 (Screening-PCR today, was run two times)<br>64.1.4 pSB1C3-CP29 (Screening-PCR today, was run two times)<br>65.1.1 pSB1C3-CP30 (Screening-PCR today, was run two times)<br>65.1.2 pSB1C3-CP30 (Screening-PCR today, was run two times)<br>65.2.1 pSB1C3-CP30 (Screening-PCR today, was run two times)<br>65.2.2 pSB1C3-CP30 (Screening-PCR today, was run two times)<br>67.1.1 pSB1C3-CP44 (Screening-PCR today, was run two times)<br>67.1.2 pSB1C3-CP44 (Screening-PCR today, was run two times)<br>67.2.1 pSB1C3-CP44 (Screening-PCR today, was run two times)<br>67.2.2 pSB1C3-CP44 (Screening-PCR today, was run two times)<br>1. pSB1C3-red (digest used in transformation, cut with E and S)<br>7. pSB3K3-red (digest used in transformation, cut with E and P)<br>22. pSB1C3-B0032-BFP (digest used in transformation, cut with X and P)<br>42. pSB4S15 (digest used in transformation, cut with SalI and SacI)<br>pSB3C17-BFP (digest used in transformation, cut with SalI and SacI)<br><br><b>PCR purification: </b><br>13.2.2 Lc. lactis MG1363-pJP059<br><br><b>Re-streaks: </b><br>42. pSB4S15-red<br>61. pSB1C3-CP1, clone 1-8 (spread out 2/7)<br>62. pSB1C3-CP8, clone 4-8 (spread out yesterday)<br>63. pSB1C3-CP11, clone 4-8 (spread out yesterday)<br>64. pSB1C3-CP29, clone 4-8 (spread out yesterday)<br>65. pSB1C3-CP30, clone 4-8 (spread out yesterday)<br>66. pSB1C3-CP41, clone 1-8 (spread out 2/7)<br>67. pSB1C3-Cp44, clone 4-8 (spread out yesterday)<br>68. pSB3K3-CP1-B0032-BFP<br>69.pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>Lb1. reuteri CF48 pLR581<br>Lb2. reuteri 1063 pLUL631<br>Lb3. reuteri DSM 20016 pVS2<br>Lb4. reuteri 100-23 no plasmid<br>Lb6. reuteri DSM 20016 pLUL63TsA8<br>Lb7. reuteri DSM 20016 pGFP<br>Lb8. reuteri 100-23 pGT232<br>Lb12. reuteri DSM 20016 no plasmid<br>Lb14. reuteri DSM 20016 pLUL631(B?)<br>Lb5. plantarum 256 rifR pAMβ1<br>Lb10. plantarum 256 no plasmid<br>Lb11. plantarum 36E no plasmid<br>43. pSB1C3-B0034-TAL<br>44. pSB1C3-B0034-His-TAL<br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br><br><b>Screening PCR and gel analysis:</b><br>45. pSB1C3-B0034-4CL<br>46. pSB1C3-B0034-His-4CL<br>47. pSB1C3-B0034-STS<br>48. pSB1C3-B0034-His-STS<br><br><b>Overnight culture:</b><br>48. pSB1C3-B0034-His-STS<br>42. pSB4S15-red<br><br><b>Gel</b> of Crt Z<br>Ligation of 38(religation of digest), 27(religation of digest), 51(religation of digest), 75, 76, 78<br>Transformation of 38(religated), 38, 27(religated), 27, 51(religated), 51, 75, 76, 78<br>O/N of 57. and 58.<br><br><h2>Results</h2><br><b>Screening PCR and gel analysis:</b><br>48.1 pSB1C3-B0034-His-STS, was successful<br>45. pSB1C3-B0034-4CL<br>47. pSB1C3-B0034-STS, failed.<br><br><b>Gel</b> of CrtZ was blank. PCR fail. Think up a new method. <br>Transformation of 75, 76 and 78(from yesterday) failed. Redo with negative controll of both ligation(by religating the digested pEL-plasmids) and plasmids(from plasmidprepp).<br>Negative controll of O/N 57. and 58. contaminated.<br><br><b>O/N:</b> <br>Lb10.1. plantarum 256: NC ok, but weirdly looking pellets and supernatants. <br>Lb11.1. plantarum 36E: NC ok, but weirdly looking pellets and supernatants. <br>Lb12.1 reuteri DSM 20016: NC ok, but weirdly looking pellets and supernatants. <br>→ Look in microscope to see if contaminated. <br><br><b>Assembly and transformation:</b> <br>1. pSB1C3-red, as NC: Good growth<br>7. pSB3K3-red, as NC: No growth<br>42. pSB4S15-red, as NC: No growth<br>61. pSB1C3-CP1: Growth, discernable red and white colonies<br>62. pSB1C3-CP8: Growth, discernable red and white colonies<br>63. pSB1C3-CP11: Growth, discernable red and white colonies<br>64. pSB1C3-CP29: Growth, discernable red and white colonies<br>65. pSB1C3-CP30: Growth, discernable red and white colonies<br>66. pSB1C3-CP41: Growth, discernable red and white colonies<br>67. pSB1C3-Cp44: Growth, discernable red and white colonies<br>77 pSB4C15-red: Growth, but only white and blue colonies*<br>*E. coli with the right plasmid should have become red.<br>→ Most seem ok, do restreaks! Test fluorescence of BFP-colonies from 2/7, get new 42’s from Erik Gullberg since not even our frozen stocks are red (as they should be).<br><br><b>Gel electrophoresis:</b> <br>Lb13.2.2 lactis MG1363 pJP059 (yesterday) No bands<br>Lb13.2.3 lactis MG1363 pJP059 (yesterday) --”--<br>Lb13.3 lactis MG1363 pJP059 (yesterday) --”--<br>Lb13.4 Lc. lactis MG1363 pJP059 (yesterday) --”--<br>Lb13.2.2 Lc. lactis MG1363 pJP059 (Touchdown) Right band (~2000)<br>Lb13.2.3 Lc. lactis MG1363 pJP059 (Touchdown) No bands<br>Lb13.3 Lc. lactis MG1363 pJP059 (Touchdown) --”--<br>Lb13.4 Lc. lactis MG1363 pJP059 (Touchdown) --”--<br>62.1 pSB1C3-CP8 (Screening) * (see below)<br>62.2 pSB1C3-CP8 (Screening) * (see below)<br>63.1 pSB1C3-CP11 (Screening) * (see below)<br>63.2 pSB1C3-CP11 (Screening) * (see below)<br>63.3 pSB1C3-CP11 (Screening) * (see below)<br>63.4 pSB1C3-CP11 (Screening) * (see below)<br>64.1.1 pSB1C3-CP29 (Screening) * (see below)<br>64.1.2 pSB1C3-CP29 (Screening) * (see below)<br>64.1.3 pSB1C3-CP29 (Screening) * (see below)<br>64.1.4 pSB1C3-CP29 (Screening) * (see below)<br>65.1.1 pSB1C3-CP30 (Screening) * (see below)<br>65.1.2 pSB1C3-CP30 (Screening) * (see below)<br>65.2.1 pSB1C3-CP30 (Screening) * (see below)<br>65.2.2 pSB1C3-CP30 (Screening) * (see below)<br>67.1.1 pSB1C3-CP44 (Screening) * (see below)<br>67.1.2 pSB1C3-CP44 (Screening) * (see below)<br>67.2.1 pSB1C3-CP44 (Screening) * (see below)<br>67.2.2 pSB1C3-CP44 (Screening) * (see below)<br>1. pSB1C3-red (cut with E and S): Clear bands (safe to use)<br>7. pSB3K3-red (cut with E and P): Clear bands (safe to use)<br>22. pSB1C3-B0032-BFP (cut with X and P) No bands (get more data)<br>42. pSB4S15 (cut with SalI and SacI) No bands (get more data)<br>pSB3C17-BFP (cut with SalI and SacI) Faint bands (likely safe)<br><br>*Neither of the two runs on gel showed anything but weak, smudgy bands. Try again.<br><br><b>PCR purification: </b><br>Lb13.2.2 Lc. lactis MG1363-pJP059: 25.9 ng/µl<br><br><b>Plasmid preparation: </b><br>Lb9.7.1 faecalis JH2-2 pAMβ1 clone 7.1 (erythromycin): 140.5 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br>Lb9.7.2 faecalis JH2-2 pAMβ1 clone 7.2 (erythromycin): 116.0 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br>Lb9.8.1 faecalis JH2-2 pAMβ1 clone 8.1 (no-antibiotic): 76.6 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br>Lb9.8.2 faecalis JH2-2 pAMβ1 clone 8.2 (no-antibiotic): 55.5 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br>Lb9.9.1 faecalis JH2-2 pAMβ1 clone 9.1 (erythromycin): 231.9 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br>Lb9.9.2 faecalis JH2-2 pAMβ1 clone 9.2 (erythromycin): 96.5 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br>Lb9.10.1 faecalis JH2-2 pAMβ1 clone 10.1 (no-antibiotic): 113.2 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br>Lb9.10.2 faecalis JH2-2 pAMβ1 clone 10.2 (no-antibiotic): 107.5 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br>Lb13.5. Lc. lactis MG1363 pJP059 clone 5: 13.8 ng/µl → Dispose & don’t save in freezer. <br>Lb13.6. Lc. lactis MG1363 pJP059 clone 6: 34.8 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br>Lb13.7. Lc. lactis MG1363 pJP059 clone 7: 25.9 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br>Lb13.8. Lc. lactis MG1363 pJP059 clone 8: 34.3 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br>Lb15.5. Lc. lactis MG1363 pGus clone 5: 5.9 ng/µl → Dispose & don’t save in freezer. <br>Lb15.6. Lc. lactis MG1363 pGus clone 6: 9.7 ng/µl → Dispose & don’t save in freezer. <br>Lb15.7. Lc. lactis MG1363 pGus clone 7: 15.7 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br>Lb15.8. Lc. lactis MG1363 pGus clone 8: 16.7 ng/µl → Throw away other low concentration clones and replace with this one in the -80o C freezer.<br><br><h2>Other experiments</h2>TBE-mix<br>Agarose gel mix<br>EDTA mix<br><br><b>Microscoping:</b> *To see if the overnights from yesterday on strains 10 → 12 were contaminated.<br>They showed to be that and the source are thought to be from a time ago, before frozen stock, since NC checked good for these overnights. </div>'; | ||
| + | } | ||
| + | else if(id == 'd2013722') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-07-22</h1><br><b>Name of participants:</b> Hampus.E, Karl H, Ken B, Kristoffer L, Victor S, Marcus H, Theodor L., Nils, Christoffer, Malin, Magnus, Niclas, Pontus, Christoffer Ahlström (CA), Stephanie Herman (SH), Alona Nyberg (AN), Mikael Strandgren (MS)<br><br><h2>Ongoing constructs</h2><br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>121.pSB1C3-B0034-STS<br>121.pSB1C3-B0034-His-STS<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>57.pSB1C3-B0034-Idi<br>99. pSB1C3-CrtO<br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>127. pSB1A3-B0034-UGTCs2-His<br>21. CrtE~I /w RBS, Zinc finger (PCR prod)<br>128. pSIM5-tet(Nissle)<br>130. pSB1C3-CrtE~I /w RBS, Zinc Finger<br>131. pSB4A15-J23101-CrtE~I /w RBS, Zinc Finger<br>132. pSB4A15-J23106-CrtE~I /w RBS, Zinc Finger<br>133. pSB4A15 -J23110-CrtE~I /w RBS, Zinc Finger<br>75 . pEL3A15 - CrtB (LR) <br>76.pEL3K16-CrtI (LR) <br>79.pEL3K16- CrtE (LR) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pSB3T5-J23101-B0034-UGTCs2-6xHis<br>142.pSB3T5-CP1-B0034-UGTCs2-6xHis<br>143.pSB3T5-CP8-B0034-UGTCs2-6xHis<br>144.pSB1C3-B0034-UGTCs2-His<br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>85. pSB4S15-CP25<br>84. pSB4S15-CP6<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>134.1 pSB1C3-CBP<br><br><h2>Todays work</h2><br><b>Restreaks:</b><br>115. pSB3T5-J23101-B0034-His-TAL*<br>116. pSB3T5-J23110-B0034-His-TAL*<br>117. pSB3T5-CP1-B0034-His-TAL*<br>137. pSB3K3-CP8-B0034-TAL*<br>138. pSB3K3-CP8-B0034-His-TAL*<br>139. pSB3K3-J23101-B0034-BFP*<br><i>-*Restreaks from the transformation, that was done the day before</i><br><br><b>Mutagenisis:</b><br>57.pSB1C3-B0034-Idi -> 129. pSB1C3-B0034-Idi (mut)<br>136.1 pSB1C3-usp45<br>136.2 pSB1C3-usp45<br>136.3 pSB1C3-usp45<br><i>→ Run PCR-products on gel to see if correct and then PCR clean it and prepare for sequencing. </i><br><br><b>Phosphorylation of primers: </b><br>usp45_mut_R<br>usp45_mut_F<br><br><b>Overnight:</b><br>127.1.pSB1A3-B0034-UGTCs2-His <br>127.2.pSB1A3-B0034-UGTCs2-His <br>86. pSB4S15-CP6-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15- CP11-B0032-BFP<br>134.1 pSB1C3-CBP<br>134.2 pSB1C3-CBP<br>134.3 pSB1C3-CBP<br>134.4 pSB1C3-CBP<br><br><b>Transformation: </b><br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>81. pSB4S15-CP41-B0032-BFP<br>82. pSB4S15-CP29-B0032-BFP<br>83. pSB4S15-CP30-B0032-BFP<br>85. pSB4S15-CP25<br><br><b>Fluorescence(BFP) experiment with UV:</b><br>84. pSB4S15-CP6<br>86. pSB4S15-CP6-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br><br><b>Screening:</b><br>131. pSB4A15-BBa-J61002-J23106-CrtE~I /w RBS, Zinc finger <br>132. pSB4A15-BBa-J61002-J23110-CrtE~I /w RBS, Zinc finger <br>133. pSB4A15-BBa-J61002-J23101-CrtE~I /w RBS, Zinc finger<br>75. pEL3A15 - CrtB <br>76. pEL3K16- CrtI <br>79. pEL3K16 - CrtE <br><br><b>Digest</b><br>3.pSB3T5-red<br>92.BBa_J61002-J23110<br>90.BBa_J61002-J23101<br>127.pSB1A3-B0034-UGTCs2-His<br>51.pEL3C18-CP1<br>51.pEL3C18-CP8<br>58.pSB1C3-B0034-IspA<br>11.pSB4A15-red<br>91.BBa_J61002-J23106<br>94.pSB1C3-K206000<br>1.pSB1C3-red<br>CSP_3<br>Plasmid 1.1 (from strain database 2013)<br><br><b>Ligation:</b><br>3.pSB3T5-red, 127.pSB1A3-B0034-UGTCs2-His, 92.BBa_J61002-J23110 --> 140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>3.pSB3T5-red, 127.pSB1A3-B0034-UGTCs2-His, 92.BBa_J61002-J23101 --> 141.pSB3T5-J23101-B0034-UGTCs2-6xHis<br>3.pSB3T5-red, 127.pSB1A3-B0034-UGTCs2-His, 51.pEL3C18-CP1 --> 142.pSB3T5-CP1-B0034-UGTCs2-6xHis<br>3.pSB3T5-red, 127.pSB1A3-B0034-UGTCs2-His, 51.pEL3C18-CP8 --> 143.pSB3T5-CP8-B0034-UGTCs2-6xHis<br>58.pSB1C3-B0034-IspA, 11.pSB4A15-red, 91.BBa_J61002-J23106 --> 108.pSBA15-J23106-B0034-IspA<br>58.pSB1C3-B0034-IspA, 11.pSB4A15-red, 94.pSB1C3-K206000 --> 109.pSBA15-K206000-B0034-IspA<br>58.pSB1C3-B0034-IspA, 11.pSB4A15-red, 95.pSB1C3-K206001 --> 110.pSBA15-K206001-B0034-IspA<br>127.pSB1A3-B0034-UGTCs2-His, 1.pSB1C3-red --> 144.pSB1C3-B0034-UGTCs2-His<br>CSP_3 <i>-Ligated with strain 1.1 (from strain database 2013) <br> Let ligate in room temperature overnight.</i> <br> <br><b>Frozen stock: (plasmid prep. tomorrow)</b><br>111.6 pSBLb4C15<br>111.7 pSBLb4C15<br><br><h2>Results</h2><br><b>Gel electrophoresis: </b><br>79.pEL3K16 - CrtE looked ok on gel, but there was primer dimer contamination. <br>75.pEL3A15 - CrtB had several bands around 1500-3000. <br>76.pEL3K16-CrtI displayed no band except primer dimers. <br>Digests looks good on gel<br><br><b>Overnights </b><br>127.1* pSB1A3-B0034-UGTCs2-His<br>127.2* pSB1A3-B0034-UGTCs2-His<br><i>-*did not grow.</i></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013723') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-07-23</h1><br><b>Name of participants:</b> Nils, Christoffer, Malin, Magnus, Niclas, Pontus, Hampus.E, Karl H, Ken B, Kristoffer L, Victor S, Marcus H, Theodor L., Christoffer Ahlström (CA), Nafisa Bashir (NB), Mikael Strandgren (MS), Stephanie Herman (SH), Alona Nyberg (AN)<br><br><h2>Ongoing constructs:</h2><br>129. pSB1C3-B0034-Idi (mut)<br>108. pSB4A15-J23106-B0034-IspA<br>109. pSB4A15-K206000-B0034-IspA<br>110. pSB4A15-K206001-B0034-IspA<br>127. pSB1A3-B0034-UGTCs2-His<br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pSB3T5-J23101-B0034-UGTCs2-6xHis<br>142.pSB3T5-CP1-B0034-UGTCs2-6xHis<br>143.pSB3T5-CP8-B0034-UGTCs2-6xHis<br>144.pSB1C3-B0034-UGTCs2-His<br>147. pSB1C3-B0034-Thioredoxin<br>152. pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br>75.pEL3A15 - CrtB (LR) <br>76.pEL3K16-CrtI (LR) <br>79.pEL3K16 - CrtE (LR) <br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>121.pSB1C3-B0034-STS<br>121.pSB1C3-B0034-His-STS<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>134. pSB1C3-CBP<br>111. pSBLb4C15<br>86. pSB4S15-CP6-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>136. pSB1C3-usp45<br><br>Lb9. faecalis JH2-2-pAMβ1<br><br><h2>Todays work</h2><br><b>Plasmid preparation:</b><br>86. pSB4S15-CP6-B0032-BFP<br>97. pSB4S15-CP8-B0032-BFP<br>98. pSB4S15-CP11-B0032-BFP<br>111.6 pSBLb4C15<br>111.7 pSBLb4C15<br>134.1 pSB1C3-CBP<br>134.2 pSB1C3-CBP<br>134.3 pSB1C3-CBP<br>134.4 pSB1C3-CBP<br>84. pSB4S15-CP6<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br><br><br><b>Mutagenesis:</b><br>136.1 pSB1C3-usp45<br>136.2 pSB1C3-usp45<br>136.3 pSB1C3-usp45<br>136.4 pSB1C3-usp45<br><br><b>PCR:</b><br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>Lb9. faecalis JH2-2-pAMβ1 clone 10.1*<br>Lb9. faecalis JH2-2-pAMβ1 clone 10.2*<br><i>-*With primers pAM-M1_F, pAM-s-X1_R and pAM-d-M1_R. To isolate ori.</i><br><br><b>Digest:</b><br>B0034-Thioredoxin, 1. pSB1C3-red (E, P) -> 147. pSB1C3-B0034-Thioredoxin<br><br><b>Ligation:</b><br>pSB1C3-B0034-Thioredoxin<br><br><b>Restreak: </b><br>139. pSB3K3-J23101-B0032-BFP<br><br><b>Transformation:</b><br>108.pSBA15-J23106-B0034-IspA<br>109.pSBA15-K206000-B0034-IspA<br>110.pSBA15-K206001-B0034-IspA<br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pSB3T5-J23101-B0034-UGTCs2-6xHis<br>142.pSB3T5-CP1-B0034-UGTCs2-6xHis<br>143.pSB3T5-CP8-B0034-UGTCs2-6xHis<br>144.pSB1C3-B0034-UGTCs2-His<br>152. pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br>135. pSB1C3-CSP*<br>148. pSB1C3-gfas purple<br>149. pSB1C3-meffred<br>150. pSB1C3-spispink<br>151. pSB1C3-meffblue<br><i>-*Transformed into D5(alpha) competent cells. </i><br><br><b>Gel electrophoresis: </b><br>136.1 pSB1C3-usp45<br>136.2 pSB1C3-usp45<br>136.3 pSB1C3-usp45<br><br><b>Screening:</b><br>129. pSB1C3-B0034-Idi (mut)<br>75.pEL3A15 - CrtE <br>76.pEL3K16 - CrtI <br>79.pEL3K16 - CrtB <br><br><b>Overnight:</b><br>127.1.pSB1A3-B0034-UGTCs2-His <br>127.2.pSB1A3-B0034-UGTCs2-His<br>84. pSB4S15-CP6<br>87. pSB4S15-CP25-B0032-BFP<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br><br><b>Spreading of strains:</b><br>135. pSB1C3-CSP*<br><i>-*Two 1x dilutions were performed.</i><br><br><b>Phosphorylation of primers: </b><br>usp45_mut_R<br>usp45_mut_F<br><br><b>Frozen stock: </b><br>134.1 pSB1C3-CBP<br>134.2 pSB1C3-CBP<br>134.3 pSB1C3-CBP<br>134.4 pSB1C3-CBP<br><br><h2>Results</h2><br><b>Previous day: </b><br><b>O/N: </b><br>134.1 pSB1C3-CBP: NC ok!<br>134.2 pSB1C3-CBP: NC ok!<br>134.3 pSB1C3-CBP: NC ok!<br>134.4 pSB1C3-CBP: NC ok!<br><br><b>Today:</b><br><b>Gel electrophoresis: </b><br>136.1 pSB1C3-usp45: Possible primer-dimer!<br>136.2 pSB1C3-usp45: Possible primer-dimer!<br>136.3 pSB1C3-usp45: Possible primer-dimer!<br><i>→ Re-do PCR mutagenesis with some edits.</i><br>129. pSB1C3-B0034-Idi (mut) did not show on gel<br>76.pEL3K16-CrtI displayed no band on gel. <br>75.pEL3A15 - CrtB displayed a band, lager than expected. <br>79.pEL3K16 - CrtE displayed a band at the right size (screened without dmso) and a good band with some contamination when screened with 7% dmso. <br><br><b>Plasmid preparation: </b><br>134.1 pSB1C3-CBP: 126.0 ng/µl → Do a sequencing preparation.<br>134.2 pSB1C3-CBP: 124.9 ng/µl<br>134.3 pSB1C3-CBP: 126.6 ng/µl<br>134.4 pSB1C3-CBP: 126.2 ng/µl<br>111.6 pSBLb4C15: 33.7 ng/µl<br>111.7 pSBLb4C15: 52.2 ng/µl<br><br><b>Transformation: </b><br>109.pSBA15-K206000-B0034-IspA - did not grow<br>147. pSB1C3-B0034-Thioredoxin - did not grow<br><br><h2>Other experiments</h2><br>10x TBE-Buffer<br>10% Glycerol solution<br><br><b>Sequencing preparation:</b><br>111.6. pSBLb4C15-red<br>111.7. pSBLb4C15-red<br>134.1 pSB1C3-CBP<br><br><b>Primer dilution (from 100 µM to 5 µM):</b><br>Ori-scr_F<br>pL1-scr_F2<br>pL1-scr_F3<br>pL3-scr_F2<br>pL3-scr_F3<br>pAM-M1_F<br>pAM-s-X1_R<br>pAM-d-M1_R</div>'; | ||
| + | } | ||
| + | else if(id == 'd2013724') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-07-24</h1><br><b>Name of participants:</b> Christoffer Ahlström (CA), Stephanie Herman (SH), Alona Nyberg (AN), Nafisa Bashir (NB), Mikael Strandgren (MS), Jens Isaksson (JI), Malin, Magnus, Pontus, Nils, Niclas, Hampus.E, Karl H, Ken B, Kristoffer L, Victor S, Marcus H.<br><br><h2>Ongoing constructs:</h2><br>127.1.pSB1A3-B0034-UGTCs2-His <br>127.2.pSB1A3-B0034-UGTCs2-His<br>108. pSBA15-J23106-B0034-IspA<br>110. pSBA15-K206001-B0034-IspA<br>140. pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141. pSB3T5-J23101-B0034-UGTCs2-6xHis<br>142. pSB3T5-CP1-B0034-UGTCs2-6xHis<br>143. pSB3T5-CP8-B0034-UGTCs2-6xHis<br>144. pSB1C3-B0034-UGTCs2-His<br>152. pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br>163.pSB3T5-ZCD-His-B0034-UGTCs2-His<br>165.pSB1C3-tsPurple<br>166.pSB1C3-scOrange<br>167.pSB1C3-fwYellow<br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>90. BBa_J61002-J23101<br>153. pSB1C3-J23110-meffBlue<br>154. pSB1C3-J23106-meffBlue<br>155. pSB1C3-J23110-meffRed<br>156. pSB1C3-J23106-meffRed<br>157. pSB1C3-J23110-spisPink<br>158. pSB1C3-J23106-spisPink<br>159. pSB1C3-J23110-gfasPurple<br>160. pSB1C3-J23106-gfasPurple<br>66. pSB1C3-CP41<br>98. pSB4S15-CP11-B0032-BFP<br>22. pSB1C3-B0032-BFP<br>87. pSB4S15-CP25-B0032-BFP<br>84. pSB4S15-CP6<br>88. pSB4S15-CP1-B0032-BFP<br>89. pSB4S15-CP44-B0032-BFP<br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119. pSB1C3-B0034-4CL<br>120. pSB1C3-B0034-HIS-4CL<br>121. pSB1C3-B0034-STS<br>121. pSB1C3-B0034-His-STS<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br><br><h2>Todays work</h2><br><b>Overnight culture: </b><br>68. pSB3K3-CP1-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br><br><b>Ligation:</b><br>139. pSB3K3-J23101-B0032-BFP<br>162. pSB1C3-J23101<br>B0034-Thioredoxin, 1. pSB1C3-red (E, P) -> 147. pSB1C3-B0034-Thioredoxin<br><br>125.1.pSB1C3-ZCD-His, 127.4.pSB1A3- B0034-UGTCs2-His -> 163.pSB3T5-ZCD-His-B0034-UGTCs2-His<br><br><b>Lambda Red:</b><br>164.E-coli Nissle-CrtY<br><br><b>Transformation: </b><br>90. BBa_J61002-J23101<br>153. pSB1C3-J23110-meffBlue<br>154. pSB1C3-J23106-meffBlue<br>155. pSB1C3-J23110-meffRed<br>156. pSB1C3-J23106-meffRed<br>157. pSB1C3-J23110-spisPink<br>158. pSB1C3-J23106-spisPink<br>159. pSB1C3-J23110-gfasPurple<br>160. pSB1C3-J23106-gfasPurple<br>147. pSB1C3-B0034-Thioredoxin<br>165.pSB1C3-tsPurple<br>166.pSB1C3-scOrange<br>167.pSB1C3-fwYellow<br><br><b>Sequencing preparation: </b><br>66. pSB1C3-CP41<br>98. pSB4S15-CP11-B0032-BFP<br>127.1.pSB1A3-B0034-UGTCs2-His <br><br><b>Digestion: </b><br>22. pSB1C3-B0032-BFP<br>90. BBa_J61002(amp)-J23101<br>125.1 pSB1C3-ZCD-His<br><br><b>Gel electrophoresis: </b><br>Lb9. faecalis JH2-2 pAMβ1<br>136.1 pSB1C3-usp45<br>136.2 pSB1C3-usp45<br>136.3 pSB1C3-usp45<br>136.4 pSB1C3-usp45<br><br><b>Re-streak:</b><br>108. pSBA15-J23106-B0034-IspA<br>110. pSBA15-K206001-B0034-IspA<br>140. pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141. pSB3T5-J23101-B0034-UGTCs2-6xHis<br>142. pSB3T5-CP1-B0034-UGTCs2-6xHis<br>143. pSB3T5-CP8-B0034-UGTCs2-6xHis<br>144. pSB1C3-B0034-UGTCs2-His<br>152. pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His <br>135.1 pSB1C3-CSP<br>135.2 pSB1C3-CSP<br>135.3 pSB1C3-CSP<br>135.4 pSB1C3-CSP<br><br><b>Mutagenesis: </b><br>136.1 pSB1C3-usp45<br>136.2 pSB1C3-usp45<br>136.3 pSB1C3-usp45<br>136.4 pSB1C3-usp45<br><br><b>PCR: </b><br>Lb9. faecalis JH2-2-pAMβ1 clone 9.9.1<br>Lb9. faecalis JH2-2-pAMβ1 clone 9.9.2<br><i>-New temperatures (-3*C), new polymerase and new primer dilutions. With primers pAM-M1_F, pAM-s-X1_R and pAM-d-M1_R. To isolate ori.</i><br><br><b>Plasmid Prep: </b><br>84.1 pSB4S15-CP6<br>84.2 pSB4S15-CP6<br>87.1 pSB4S15-CP25-B0032-BFP<br>87.2 pSB4S15-CP25-B0032-BFP<br>88.1 pSB4S15-CP1-B0032-BFP<br>88.2 pSB4S15-CP1-B0032-BFP<br>89.1 pSB4S15-CP44-B0032-BFP<br>89.2 pSB4S15-CP44-B0032-BFP<br>127.1.pSB1A3-B0034-UGTCs2-His <br>127.2.pSB1A3-B0034-UGTCs2-His<br><br><b>Screening PCR and gel electrophoresis on:</b><br>115. pSB3T5-J23101-B0034-His-TAL clone: 1, 2, 3 ,4<br>116. pSB3T5-J23110-B0034-His-TAL clone: 1, 2, 3, 4<br>117. pSB3T5-CP1-B0034-His-TAL clone: 1, 2, 3, 4<br>137. pSB3K3-CP8-B0034-TAL clone: 1, 2, 3, 4<br>138. pSB3K3-CP8-B0034-His-TAL clone: 1, 2, 3, 4<br>139. pSB3K3-J23101-B0034-BFP clone: 1, 2, 3, 4<br>Sequence prep on: <br>116. pSB3T5-J23110-B0034-His-TAL clone: 3, 4<br>137. pSB3K3-CP8-B0034-TAL clone: 1, 4<br>Overnight culture on: <br>116. pSB3T5-J23110-B0034-His-TAL clone: 3, 4<br>137. pSB3K3-CP8-B0034-TAL: 1, 4<br>Measurement of standards of p-coumaric acid in the shimadzu UV160, plotting of a standard graph.<br><br><h2>Results</h2><br><b>Previous day:</b><br><b>Spreading of strains: </b><br>135. pSB1C3-CSP: Both white and red colonies.<br><i>*Two 1x dilutions were performed.<br>→ Re-streak with 4 colonies.</i><br><br><b>PCR:</b> <br>One of the PCR-machines (with 2x MluI-pAM) <br><i>-were accidentally paused right after start last night, and so it had to be re-done this morning. Since the samples stood in room temperature for about 15 hour the polymerase may have lost it’s activity, a gel will tell.</i><br><br><b>Today:</b><br><b>Gel electrophoresis: </b><br>Lb9.10.1 faecalis JH2-2 pAMβ1 with XbaI and MluI: Not even primer-bands<br>Lb9.10.2 faecalis JH2-2 pAMβ1 with 2x MluI: -”-<br>Lb9.10.1 faecalis JH2-2 pAMβ1 with XbaI and MluI: -”-<br>Lb9.10.2 faecalis JH2-2 pAMβ1 with 2x MluI: -”-<br>136.1 pSB1C3-usp45: Possible primer-dimer and a mark at ~1000bp.<br>136.2 pSB1C3-usp45: Possible primer-dimer.<br>136.3 pSB1C3-usp45: Possible primer-dimer.<br>136.4 pSB1C3-usp45: Possible primer-dimer.<br><i>→ Re-do PCR with other temperatures!</i><br><br><b>Plasmid Prepp: </b><br>84.1 pSB4S15-CP6: 59.3 ng/µl<br>84.2 pSB4S15-CP6: 66.7 ng/µl<br>87.1 pSB4S15-CP25-B0032-BFP: 12.6 ng/µl <br>87.2 pSB4S15-CP25-B0032-BFP: 52.0 ng/µl<br>88.1 pSB4S15-CP1-B0032-BFP: 508.2 ng/µl <br>88.2 pSB4S15-CP1-B0032-BFP: 489.7 ng/µl<br>89. pSB4S15-CP44-B0032-BFP: 7.4 ng/µl<br><br><b>digest</b> of B0034-Thioredoxin (Synthetic construct) looks good on gel<br><br><b>Restreak</b> of 143. pSB3T5-CP8-B0034-UGTCs2-6xHis did not grow<br><br><b>ligation</b> of 141. pSB3T5-J23101-B0034-UGTCs2-6xHis is wrong, since 92.BBa_J61002-J23101 was digested wrong<br><br>Construct 116.3, 116.4, 137.1, 137.4 successfull.<br><br><h2>Other experiments</h2><br><b>Primer dilutions (from 100µM to 5 µM): </b><br>pAM-M1_F<br>pAM-M1_R<br>pAM-X1_R<br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013725') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-07-25</h1><br><b>Name of participants:</b> Malin, Magnus, Niclas, Pontus, Nils, Nafisa Bashir (NB), Viktor Blomkvist (VB), Jens Isaksson (JI), Mikael Strandgren (MS), Hampus.E, Kristoffer L, Victor S, Marcus H, Karl H, Theodor L<br><br><h2>Ongoing constructs</h2><br><b>E. coli:</b><br>108.pSBA15-J23106-B0034-IspA<br>110.pSBA15-K206001-B0034-IspA<br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pSB3T5-J23101-B0034-UGTCs2-6xHis <br>142.pSB3T5-CP1-B0034-UGTCs2-6xHis<br>143.pSB3T5-CP8-B0034-UGTCs2-6xHis<br>144.pSB1C3-B0034-UGTCs2-His<br>147.pSB1C3-B0034-Thioredoxin<br>152.pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br>163.pSB3T5-ZCD-His-B0034-UGTCs2-His<br>164.E-coli Nissle-CrtY<br>165.pSB1C3-tsPurple<br>166.pSB1C3-scOrange<br>167.pSB1C3-fwYellow<br>135. pSB1C3-CSP<br>136.1 pSB1C3-usp45<br>148. pSB1C3-gfas purple<br>149. pSB1C3-meffred<br>150. pSB1C3-spispink<br>151. pSB1C3-meffblue<br>162. pSB1C3-J23101<br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>121.pSB1C3-B0034-STS<br>121.pSB1C3-B0034-His-STS<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br><br><b>E. faecalis:</b><br>9. JH2-2-pAMβ1 clone<br><br><h2>Todays work</h2><br><b>O/N:</b> <br>135. pSB1C3-CSP<br><br><b>Transformation:</b><br>163.pSB3T5-ZCD-His-B0034-UGTCs2-His<br>143. pSB3T5-CP8-B0034-UGTCs2-6xHis<br>162. pSB1C3-J23101<br><br><b>Restreak:</b><br>139. pSB3K3-J23101-B0032-BFP<br>148. pSB1C3-gfas purple<br>149. pSB1C3-meffred<br>150. pSB1C3-spispink<br>151. pSB1C3-meffblue<br>147. pSB1C3-B0034-Thioredoxin<br>164.E-coli Nissle-CrtY<br>165.pSB1C3-tsPurple<br>166.pSB1C3-scOrange<br>167.pSB1C3-fwYellow<br><br><b>Plasmid prep on:</b><br>116. pSB3T5-J23110-B0034-His-TAL clone: 3, 4<br>137. pSB3K3-CP8-B0034-TAL clone: 1, 4<br><br><br><b>Gel electrophoresis of PCR products: </b><br>Lb9. faecalis JH2-2-pAMβ1 clone 9.9.1*<br>Lb9. faecalis JH2-2-pAMβ1 clone 9.9.2*<br>136.1 pSB1C3-usp45<br>136.2 pSB1C3-usp45<br>136.3 pSB1C3-usp45<br>136.4 pSB1C3-usp45<br><i>-*(With primers pAM-M1_F, pAM-s-X1_R and pAM-d-M1_R)</i><br><br><b>PCR-purification: </b><br>Lb9. faecalis JH2-2-pAMβ1 clone 9.9.1*<br>Lb9. faecalis JH2-2-pAMβ1 clone 9.9.2*<br><i>*With primers pAM-M1_F, pAM-s-X1_R and pAM-d-M1_R</i><br><br><b>Screening:</b><br>108.3 pSBA15-J23106-B0034-IspA<br>108.4 pSBA15-J23106-B0034-IspA<br>110.1 pSBA15-K206001-B0034-IspA<br>140.1 pSB3T5-J23110-B0034-UGTCs2-6xHis<br>140.2 pSB3T5-J23110-B0034-UGTCs2-6xHis<br>140.3 pSB3T5-J23110-B0034-UGTCs2-6xHis<br>140.4 pSB3T5-J23110-B0034-UGTCs2-6xHis<br>142.1 pSB3T5-CP1-B0034-UGTCs2-6xHis<br>142.2 pSB3T5-CP1-B0034-UGTCs2-6xHis<br>142.3 pSB3T5-CP1-B0034-UGTCs2-6xHis<br>142.4 pSB3T5-CP1-B0034-UGTCs2-6xHis<br>144.1 pSB1C3-B0034-UGTCs2-His<br>144.2 pSB1C3-B0034-UGTCs2-His<br>144.3 pSB1C3-B0034-UGTCs2-His<br>144.4 pSB1C3-B0034-UGTCs2-His<br><br><b>Overnight:</b><br>144.1 pSB1C3-B0034-UGTCs2-His<br>144.2 pSB1C3-B0034-UGTCs2-His<br>144.3 pSB1C3-B0034-UGTCs2-His<br>144.4 pSB1C3-B0034-UGTCs2-His<br>152.1 pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br>152.2 pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br>152.3 pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br>152.4 pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br><br><b>Ligation:</b><br>3.pSB3T5-red, 127.pSB1A3-B0034-UGTCs2-His, 103.pSB1K3-J23101--> 141.pSB3T5-J23101-B0034-UGTCs2-6xHis <br><br><b>Digest and assembly:</b><br>pL3E1 (pAMβ1 double)<br>pL3 (pAMβ1 single)<br>105. pSB3T5-J23101-B0034-TAL<br>115. pSB3T5-J23101-B0034--His-TAL<br><br><b>Transformation on construct:</b><br>105. pSB3T5-J23101-B0034-TAL<br>115. pSB3T5-J23101-B0034--His-TAL<br><br><h2>Results</h2><br><b>Screening of all clones failed except for:</b><br>144.1 pSB1C3-B0034-UGTCs2-His<br>144.2 pSB1C3-B0034-UGTCs2-His<br>144.3 pSB1C3-B0034-UGTCs2-His<br>144.4 pSB1C3-B0034-UGTCs2-His<br><br><b>Today:</b><br><b>Gel electrophoresis: </b><br>Lb9. faecalis JH2-2-pAMβ1 clone 9.9.1: Clear and correct bands, at ~1700 bp.*<br>Lb9. faecalis JH2-2-pAMβ1 clone 9.9.2: Clear and correct bands, at ~1700 bp.*<br>136.1 pSB1C3-usp45: Big smudge between approx. 2500 and 200 bp<br>136.2 pSB1C3-usp45: Empty<br>136.3 pSB1C3-usp45: Empty<br>136.4 pSB1C3-usp45: Empty<br> <i>*(With primers pAM-M1_F, pAM-s-X1_R and pAM-d-M1_R)</i><br><br><h2>Other experiments</h2><br>75. Digest to check for wrong bindings<br>Making LB-agar plates<br><br>Spectrophotomertry of 106.5, 106.6 and 43.7, using the shimadzu UV160.<br><i>-Inconclusive spectrophotometry results.</i></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013726') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-07-26</h1><br><b>Name of participants:</b> Nafisa Bashir (NB), Christoffer Ahlström (CA), Stephanie Herman (SH), Alona Nyberg (AN), Mikael Strandgren (MS), Viktor Blomkvist (VB), Jens Isaksson (JI), Malin, Magnus, Niclas, Pontus, Nils, Hampus.E, Kristoffer L, Viktor @, Marcus H. Ken B-A, Karl H, Theodor L<br><br><h2>Ongoing constructs</h2><br>148. pSB1C3-gfas purple<br>149. pSB1C3-meffred<br>150. pSB1C3-spispink<br>151. pSB1C3-meffblue<br>90. BBa_J61002-J23101<br>153. pSB1C3-J23110-meffBlue<br>154. pSB1C3-J23106-meffBlue<br>155. pSB1C3-J23110-meffRed<br>156. pSB1C3-J23106-meffRed<br>157. pSB1C3-J23110-spisPink<br>158. pSB1C3-J23106-spisPink<br>159. pSB1C3-J23110-gfasPurple<br>160. pSB1C3-J23106-gfasPurple<br>162. pSB1C3-J23101<br>168. pSBLbC2 (working name)<br>169. pSBLbEcC2 (working name)<br>143. pSB3T5-CP8-B0034-UGTCs2-6xHis<br>144.pSB1C3-B0034-UGTCs2-His<br>152.pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br>147. pSB1C3-B0034-Thioredoxin<br>164.E-coli Nissle-CrtY<br>165.pSB1C3-tsPurple<br>166.pSB1C3-scOrange<br>167.pSB1C3-fwYellow<br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>121.pSB1C3-B0034-STS<br>121.pSB1C3-B0034-His-STS<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br><br><h2>Todays work</h2><br>Screening:<br>108.3 pSBA15-J23106-B0034-IspA<br>108.4 pSBA15-J23106-B0034-IspA<br>110.1 pSBA15-K206001-B0034-IspA<br>140.1 pSB3T5-J23110-B0034-UGTCs2-6xHis<br>140.2 pSB3T5-J23110-B0034-UGTCs2-6xHis<br>140.3 pSB3T5-J23110-B0034-UGTCs2-6xHis<br>140.4 pSB3T5-J23110-B0034-UGTCs2-6xHis<br>142.1 pSB3T5-CP1-B0034-UGTCs2-6xHis<br>142.2 pSB3T5-CP1-B0034-UGTCs2-6xHis<br>142.3 pSB3T5-CP1-B0034-UGTCs2-6xHis<br>142.4 pSB3T5-CP1-B0034-UGTCs2-6xHis<br>147. pSB1C3-B0034-Thioredoxin<br>165.pSB1C3-tsPurple<br>166.pSB1C3-scOrange<br>167.pSB1C3-fwYellow<br><br></b>Re-streak: </b><br>163.pSB3T5-ZCD-His-B0034-UGTCs2-His<br>143. pSB3T5-CP8-B0034-UGTCs2-6xHis<br>105. pSB3T5-J23101-B0034-TAL<br>115. pSB3T5-J23101-B0034--His-TAL<br><br>Plasmid prep:<br>144.1 pSB1C3-B0034-UGTCs2-His<br>144.2 pSB1C3-B0034-UGTCs2-His<br>144.3 pSB1C3-B0034-UGTCs2-His<br>144.4 pSB1C3-B0034-UGTCs2-His<br>152.1 pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br>152.2 pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br>152.3 pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br>152.4 pUC57 (amp)-B0034-Exp-Linker-Miraculin-Linker-His<br><br><b>Transformation*:</b><br>168. pSBLbC2 (ori from pSB4C15 replaced with ori from pAMβ1)<br>169. pSBLbEcC2 (ori from pAMβ1 added to pSB4C15)<br><i>* Not spread.</i><br><br><b>Ligation:</b><br>3.pSB3T5-red, 127.pSB1A3-B0034-UGTCs2-His, 103.pSB1K3-J23101--> 141.pSB3T5-J23101-B0034-UGTCs2-6xHis <br><br><h2>Results:</h2><br>Screening showed incorrect results on all constructs, but possible sources for error in the analysis made these results inconclusive. A new screening was set to run overnight.<br><br><h2>Other experiments</h2><br>Organization of frozen stock<br><br>75. Digest to check for wrong bindings<br><br>Two phase separation of p-coumaric acid with heptane. Spectrophotometry.<br><i>--Heptane proved not to be so good for the separation.</i></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013727') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-07-27</h1><br><b>Name of participants:</b> Kristoffer L, Marcus H., Stephanie Herman (SH), Nafisa Bashir (NB), Malin, Magnus, Pontus, Nils<br><br><h2>Ongoing constructs</h2><br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>121.pSB1C3-B0034-STS<br>121.pSB1C3-B0034-His-STS<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>170. pSB3T5-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>90. BBa_J61002-J23101<br>148. pSB1C3-gfas purple<br>149. pSB1C3-meffred<br>150. pSB1C3-spispink<br>151. pSB1C3-meffblue<br>153. pSB1C3-J23110-meffBlue<br>154. pSB1C3-J23106-meffBlue<br>155. pSB1C3-J23110-meffRed<br>156. pSB1C3-J23106-meffRed<br>157. pSB1C3-J23110-spisPink<br>158. pSB1C3-J23106-spisPink<br>159. pSB1C3-J23110-gfasPurple<br>160. pSB1C3-J23106-gfasPurple<br>162. pSB1C3-J23101<br><br><h2>Todays work</h2><br><b>Digest on construct:</b><br>106. pSB3T5-J23110-B0034-TAL<br>104. pSB1C3-B0034-4CL-B0034-STS<br><br><b>Assembly on cnstruct:</b><br>170. pSB3T5-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>Transformation on construct:<br>170. pSB3T5-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br><br><b>Plasmid Preperation: </b><br>148.1 pSB1C3-gfas purple<br>148.2 pSB1C3-gfas purple<br>149.1 pSB1C3-meffred<br>149.4 pSB1C3-meffred<br>150.1 pSB1C3-spispink<br>150.2 pSB1C3-spispink<br>151.2 pSB1C3-meffblue<br><br><b>Overnight culture:</b> <br>90. BBa_J61002-J23101<br>153. pSB1C3-J23110-meffBlue<br>154. pSB1C3-J23106-meffBlue<br>155. pSB1C3-J23110-meffRed<br>156. pSB1C3-J23106-meffRed<br>157. pSB1C3-J23110-spisPink<br>158. pSB1C3-J23106-spisPink<br>159. pSB1C3-J23110-gfasPurple<br>160. pSB1C3-J23106-gfasPurple<br>162. pSB1C3-J23101<br><br><h2>Result</h2><br><b>Transformation on construct:</b><br>170. pSB3T5-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>Failed. Retry assembly<br>110.pSBA15-K206001-B0034-IspA did not grow<br><br><b>Gel electrophoresis: </b><br>147. pSB1C3-B0034-Thioredoxin<br>165.pSB1C3-tsPurple<br>166.pSB1C3-scOrange<br>167.pSB1C3-fwYellow<br>didn’t show on gel<br><br><b>Plasmid Preperation:</b><br>148.1 pSB1C3-gfas purple: 311.6 ng/µl<br>148.2 pSB1C3-gfas purple: 338.0 ng/µl<br>149.1 pSB1C3-meffred: 202.4 ng/µl<br>149.4 pSB1C3-meffred: 310.4 ng/µl<br>150.1 pSB1C3-spispink: 305.4 ng/µl<br>150.2 pSB1C3-spispink: 323.8 ng/µl<br>151.2 pSB1C3-meffblue: 179.2 ng/µl<br><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013728') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Sunday 2013-07-28</h1><br><b>Name of participants:</b> Mikael Strandgren (MS), Alona Nyberg(AN), Kristoffer L, Marcus H, Karl H.,Malin, Magnus, Nils, Pontus<br><br><h2>Ongoing constructs</h2><br><b>E. coli:</b><br>168. pSBLbC2<br>169. pSBLbEcC2<br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>121.pSB1C3-B0034-STS<br>121.pSB1C3-B0034-His-STS<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>170. pSB3T5-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>173. pSB3T5-CP1-B0034-4CL-B0034-STS<br>174. pSB3T5-CP8-B0034-4CL-B0034-STS<br>175. pSB3T5-J23101-B0034-4CL-B0034-STS<br> <br><h2>Todays work</h2><br><b>PCR:</b><br>19. CrtE~(O)-Z/w RBS, ZF<br>20.CrtE~Z/w RBS, ZF<br>21. CrtE~I/wRBS,ZF<br><br><b>Restreak:</b><br>140.pSB3T5-J23110-B0034-UGTCs2-His<br>141.pSB3T5-J23101-B0034-UGTCs2-6xHis<br>142.pSB3T5-CP1-B0034-UGTCs2-His.<br><br><b>Spreading of transformed E. coli (from 20130726):</b><br>90.1 BBa_J61002-J23101<br>153. pSB1C3-j23110-meffBlue <br>155. pSB1C3-j23110-meffRed<br>156. pSB1C3-j23106-meffRed<br>157. pSB1C3-j23110-spisPink<br>159. pSB1C3-j23110-gfasPurple<br>160. pSB1C3-j23106-gfasPurple<br>162. pSB1C3-J23101<br>168. pSBLbC2<br>169. pSBLbEcC2<br><br><b>Frozen stock & plasmid preparation:</b><br>90.1 BBa_J61002-J23101<br>90.2 BBa_J61002-J23101<br>153.1 pSB1C3-j23110-meffBlue <br>153.2 pSB1C3-j23110-meffBlue<br>155.1 pSB1C3-j23110-meffRed<br>156.1 pSB1C3-j23106-meffRed<br>157.1 pSB1C3-j23110-spisPink<br>157.2 pSB1C3-j23110-spisPink<br>159.4 pSB1C3-j23110-gfasPurple<br>160.3 pSB1C3-j23106-gfasPurple<br>160.4 pSB1C3-j23106-gfasPurple<br>162.1 pSB1C3-J23101<br>162.2 pSB1C3-J23101<br><br><b>Overnight:</b><br>106.5 pSB3K3-B0034-J23110-TAL<br>106.6 pSB3K3-B0034-J23110-TAL<br>43.7 pSB1C3-B0034-TAL<br>143.1.pSB3T5-CP8-B0034-UGTCs2-6xHis<br>167.1.pSB1C3-fwYellow<br>167.2.pSB1C3-fwYellow <br><br><b>Digest:</b><br>5.pSB4K15-red digested with E,P<br>111.6.pSBLb4C15-red digested with E,P<br>152.1.pUC57(amp)-B0034-ExpTag-Linker-Miraculin-HisTag digested with E,P<br>152.1.pUC57(amp)-B0034-ExpTag-Linker-Miraculin-HisTag digested with X,P<br><br><b>Assembly:</b><br>105. pSB3T5-J23101-B0034-TAL<br>170. pSB3T5-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>173. pSB3T5-CP1-B0034-4CL-B0034-STS<br>174. pSB3T5-CP8-B0034-4CL-B0034-STS<br>175. pSB3T5-J23101-B0034-4CL-B0034-STS<br><br><b>Ligation:</b><br>58.pSB1C3-B0034-IspA, 5.pSB4K15-red, 91.BBa_J61002-J23106 --> 108.pSBA15-J23106-B0034-IspA<br>58.pSB1C3-B0034-IspA, 11.pSB4A15-red, 94.pSB1C3-K206000--> 109.pSBA15-K206000-B0034-IspA<br>58.pSB1C3-B0034-IspA, 11.pSB4A15-red, 95.pSB1C3-K206001 --> 110.pSBA15-K206001-B0034-IspA<br>3.pSB3T5-red, 125.1.pSB1C3-ZCD-His, 127.4.pSB1A3- B0034-UGTCs2-His -> 163.pSB3T5-ZCD-His-B0034-UGTCs2-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(E,P), 1.pSB1C3-red-> 176.B1C3-B0034-Exp-Linker-Miraculin-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 3.pSB3T5-red, 103.pSB1K3-J23101 -> 177.pSB3T5-J23101-B0034-Exp-Linker-Miraculin-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 3.pSB3T5-red, 92.BBa_J61002(amp)_J23110 -> 178.pSB3T5-J23110-B0034-Exp-Linker-Miraculin-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 3.pSB3T5-red, CP1 -> 179.pSB3T5-CP1-B0034-Exp-Linker-Miraculin-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 3.pSB3T5-red, CP8 -> 180.pSB3T5-CP8-B0034-Exp-Linker-Miraculin-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 111.pSBLb4C15-red(E,P), CP1 -> 181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 111.pSBLb4C15-red(E,P), CP8 -> 182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br><br><b>Transformation:</b><br>105. pSB3T5-J23101-B0034-TAL<br>170. pSB3T5-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>173. pSB3T5-CP1-B0034-4CL-B0034-STS<br>174. pSB3T5-CP8-B0034-4CL-B0034-STS<br>175. pSB3T5-J23101-B0034-4CL-B0034-STS<br><br><b>Gel electrophoresis: </b><br>76. pEL3K16-CrtI<br>20. CrtZ<br>19.CrtE~(O)-Z<br>20.CrtE~Z<br>21. CrtE~I<br>143.pSB3T5-CP8-B0034-UGTCs2-6xHis<br>163.pSB3T5-ZCD-His-B0034-UGTCs2-His<br>147.pSB1C3-B0034-Thioredoxin<br>165.pSB1C3-tsPurple<br>166.pSB1C3-scOrange<br>167.pSB1C3-fwYellow<br><br><b>Sequence prep:</b><br>144. pSB1C3-B0034-UGTCs2-His<br><br><h2>Results</h2><br><b>Frozen stock & plasmid preparation:</b><br>90.1 BBa_J61002-J23101: 215.4 ng/µl <br>90.3 BBa_J61002-J23101: 210.1 ng/µl<br>153.1 pSB1C3-j23110-meffBlue: 112.6 ng/µl<br>153.2 pSB1C3-j23110-meffBlue: 119 ng/µl<br>155.1 pSB1C3-j23110-meffRed: 169.5 ng/µl<br>156.1 pSB1C3-j23106-meffRed: 200.3 ng/µl<br>157.1 pSB1C3-j23110-spisPink: 73.7 ng/µl<br>157.2 pSB1C3-j23110-spisPink: 222.1 ng/µl<br>159.4 pSB1C3-j23110-gfasPurple: 279.7 ng/µl<br>160.3 pSB1C3-j23106-gfasPurple: 116.1 ng/µl<br>160.4 pSB1C3-j23106-gfasPurple: 122.8 ng/µl<br>162.1 pSB1C3-J23101: 79.9 ng/µl<br>162.2 pSB1C3-J23101: 80.1 ng/µl<br><br><b>Gel electrophoresis: </b><br>76. pEL3K16-CrtI - Correct at 50 degrese tube<br>20. CrtZ - Correct for low temps<br>19.CrtE~(O)-Z - weak incorrect bands<br>20.CrtE~Z - weak incorrect bands<br>21. CrtE~I - 3 bands, one correct 2 wrong<br>143. pSB3T5-CP8-B0034-UGTCs2-6xHis - 143.1 success, other clones failed<br>163.pSB3T5-ZCD-His-B0034-UGTCs2-His - Failed<br>167.1.pSB1C3-fwYellow - Success<br>167.2.pSB1C3-fwYellow - Success<br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013729') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-07-29</h1><br><b>Name of participants:</b> Malin, Magnus, Pontus, Nils, Christoffer, Kristoffer L, Marcus H, Karl H, Theodor L, Emil M, Victor S, Alona Nyberg(AN), Nafisa Bashir (NB), Mikael Strandgren (MS), Jens Isaksson (JI)<br><br><h2>Ongoing constructs</h2><br>108.pSB4K15-J23106-B0034-IspA<br>109.pSB4A15-K206000-B0034-IspA<br>110.pSB4A15-K206001-B0034-IspA<br>163.pSB3T5-ZCD-His-B0034-UGTCs2-His<br>176.B1C3-B0034-Exp-Linker-Miraculin-His<br>177.pSB3T5-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pSB3T5-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pSB3T5-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pSB3T5-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>121.pSB1C3-B0034-STS<br>121.pSB1C3-B0034-His-STS<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>170. pSB3T5-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>173. pSB3T5-CP1-B0034-4CL-B0034-STS<br>174. pSB3T5-CP8-B0034-4CL-B0034-STS<br>175. pSB3T5-J23101-B0034-4CL-B0034-STS<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br>Lb10. plantarum 256-p256<br><br><h2>Todays work</h2><br><b>O/N, Frozen Stock:</b><br>76.pEL3K16-CrtI<br><br><b>Screening & Overnight:</b><br>140.pSB3T5-J23110-B0034-UGTCs2-His<br>141.pSB3T5-J23101-B0034-UGTCs2-6xHis<br>142.pSB3T5-CP1-B0034-UGTCs2-His.<br><br><b>Digestion: </b><br>3. pSB3K3-red<br>36. pEL3S17-red<br>106. pSB3T5-J23110-B0034-TAL<br><br><b>Assembly:</b><br>170. pSB3T5-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>173. pSB3T5-CP1-B0034-4CL-B0034-STS<br>174. pSB3T5-CP8-B0034-4CL-B0034-STS<br>175. pSB3T5-J23101-B0034-4CL-B0034-STS<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br><br><b>Transformation:</b><br>108.pSBA15-J23106-B0034-IspA<br>109.pSBA15-K206000-B0034-IspA<br>110.pSBA15-K206001-B0034-IspA<br>163.pSB3T5-ZCD-His-B0034-UGTCs2-His<br>176.B1C3-B0034-Exp-Linker-Miraculin-His<br>177.pSB3T5-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pSB3T5-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pSB3T5-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pSB3T5-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>170. pSB3T5-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>173. pSB3T5-CP1-B0034-4CL-B0034-STS<br>174. pSB3T5-CP8-B0034-4CL-B0034-STS<br>175. pSB3T5-J23101-B0034-4CL-B0034-STS<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br>Lb10. plantarum 256 p256 (as competence test)<br>Lb11. plantarum 256 plasmid free (as positive control)<br>Lb12. reuteri DSM-20016 plasmid free (as positive control)<br>Lb23. reuteri 100-23-pSBLb4C15-red (single-ori (pJP059) shuttle vector)<br>Lb24. reuteri DSM-20016-pSBLb4C15-red (--”--)<br>Lb25. reuteri 100-23-pSBLbC2-red (single-ori (pAMbeta1) shuttle vector)<br>Lb28. reuteri 100-23-pVS2-red (as competence test)<br><br><b>Restreak:</b><br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>173. pSB3T5-CP1-B0034-4CL-B0034-STS<br>174. pSB3T5-CP8-B0034-4CL-B0034-STS<br>175. pSB3T5-J23101-B0034-4CL-B0034-STS<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br><br><b>Ligation:</b><br>120. pSB1C3-B0034-His-4CL<br>173. pSB3T5-CP1-B0034-4CL-B0034-STS<br><br><b>Gel:</b><br>147.pSB1C3-B0034-Thioredoxin<br>165.pSB1C3-tsPurple<br>166.pSB1C3-scOrange<br>167.pSB1C3-fwYellow<br>19. CrtE~(O)-Z/w RBS, ZF<br>20. CrtE~Z/w RBS, ZF + gelextraction<br>21. CrtE~I/w RBS, ZF<br>Lb10. plantarum 256 p256 <br>169. pSBLbEcC2-red (20 clones)*<br><i>-* With the purpose of trying to find clones transformed with plasmids containing the pAMbeta1-ori as well as the E. coli ori.</i><br><br><b>Digest, ligate and transformation:</b> <br>90.1 BBa_J61002-J23101 to 139. pSB3K3-J23101-B0032-BFP<br><br>Two phase separation with octanol and acetic acid. Spectrophotometry of 106.5 and 106.6. Comparison between the Shimadzu UV160, Shimadzu UV1800, and the nanodrop.<br><br><h2>Results</h2><br><b>Gel:</b><br>147.pSB1C3-B0034-Thioredoxin - success<br>165.pSB1C3-tsPurple - success<br>166.pSB1C3-scOrange - success<br>167.pSB1C3-fwYellow - success<br>19. CrtE~(O)-Z/w RBS, ZF - No right band<br>20. CrtE~Z/w RBS, ZF - Right band slightly above 10000bp, thou weak and others,<br>21. CrtE~I/w RBS, ZF - Right band at around 8000bp<br>All tranformations but 176 failed. Ligatoins will be redone, but with pSB3T5 replaced with pEL3S17, as this backbone has shown better performance.<br><br><b>Transformation succes on construct:</b><br>173. pSB3T5-CP1-B0034-4CL-B0034-STS<br>174. pSB3T5-CP8-B0034-4CL-B0034-STS<br>175. pSB3T5-J23101-B0034-4CL-B0034-STS<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br><br><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013730') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-07-30</h1><br><b>Name of participants:</b> Christoffer Ahlström (CA), Alona Nyberg (AN), Mikael Strandgren (MS), Malin, Magnus, Pontus, Nils, Christoffer, Sabri, Thorsteinn, Marcus H, Karl H, Theodor L, Emil M, Victor S<br><br><h2>Ongoing constructs:</h2><br><b>E.coli:</b><br>169. pSBLbEcC2-red<br>191. pSB1A2-fusionGFP<br>108.pSBA15-J23106-B0034-IspA<br>109.pSBA15-K206000-B0034-IspA<br>110.pSBA15-K206001-B0034-IspA<br>143.pSB3T5-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>176.pSB1C3-B0034-Exp-Linker-Miraculin-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSb1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>104. pSB1C3-B0034-4CL-B0034-STS<br>105. pSB3T5-J23101-B0034-TAL<br>106. pSB3T5-J23110-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3T5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>117. pSB3T5-CP1-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>121.pSB1C3-B0034-STS<br>121.pSB1C3-B0034-His-STS<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>170. pSB3T5-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>173. pSB3T5-CP1-B0034-4CL-B0034-STS<br>174. pSB3T5-CP8-B0034-4CL-B0034-STS<br>175. pSB3T5-J23101-B0034-4CL-B0034-STS<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br><br><h2>Todays work</h2><br><b>Transformation: </b><br>191. pSB1A2-fusionGFP<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>120. pSB1C3-B0034-His-4CL<br>Spectrofotometry (using the nanodrop) of 106.5 and 106.6. Inconclusive results.<br><br><b>Screening:</b><br>140.pSB3T5-J23110-B0034-UGTCs2-His<br>141.pSB3T5-J23101-B0034-UGTCs2-6xHis<br>142.pSB3T5-CP1-B0034-UGTCs2-His.<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS clone: 1,2,3,4<br>172. pSB3T5-CP1-B0034-TAL clone: 1,2,3,4<br><br><b>Plasmid preparation:</b><br>143.pSB3T5-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>165.pSB1C3-tsPurple<br>166.pSB1C3-scOrange<br>167.pSB1C3-fwYellow<br><br><b>PCR:</b><br>19. CrtE~(O)-Z/w RBS, ZF<br>20. CrtE~Z/w RBS, ZF<br><br><b>Phosphorylation:</b><br>20. CrtE~Z/w RBS, ZF<br>21. CrtE~I/w RBS, ZF<br><br><b>Streaking of strains: </b><br>191. pSB1A2-fusionGFP<br><br><b>Re-streak: </b><br>139. pSB3K3-J23101-B0032-BFP<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br>176.pSB1C3-B0034-Exp-Linker-Miraculin-His<br><br><b>Ligate and transformation: </b><br>90. BBa_J61002-J23101 to 139. pSB3K3-J23101-B0032-BFP<br><br><b>Digest and ligate: </b><br>162. pSB1C3-J23101 to 192.pSB3K3-J23101-B0032-BFP<br>58.pSB1C3-B0034-IspA, 5.pSB4K15-red, 91.BBa_J61002-J23106 --> 108.pSBA15-J23106-B0034-IspA<br>58.pSB1C3-B0034-IspA, 11.pSB4A15-red, 94.pSB1C3-K206000--> 109.pSBA15-K206000-B0034-IspA<br>58.pSB1C3-B0034-IspA, 11.pSB4A15-red, 95.pSB1C3-K206001 --> 110.pSBA15-K206001-B0034-IspA<br>36. pEL3S17-red, 125.1.pSB1C3-ZCD-His, 127.4.pSB1A3- B0034-UGTCs2-His -> 163. pEL3S17-ZCD-His-B0034-UGTCs2-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 36.pEL3S17-red, 103.pSB1K3-J23101 -> 177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 36.pEL3S17-red, 92.BBa_J61002(amp)_J23110 -> 178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 36.pEL3S17-red, CP1 -> 179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 36.pEL3S17-red, CP8 -> 180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 111.pSBLb4C15-red(E,P), CP1 -> 181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 111.pSBLb4C15-red(E,P), CP8 -> 182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br><br><b>O/N, Frozen Stock:</b><br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSb1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br><br><b>Gel electrophoresis on:</b><br>171. pSB3T5-J23110-B0034-4CL-B0034-STS clone: 1,2,3,4<br>172. pSB3T5-CP1-B0034-TAL clone: 1,2,3,4<br><br><h2>Results</h2><br><b>Previous day: </b><br><br><b>Transformation:</b><br>Lb10. plantarum 256 p256 growing, possibly contaminated<br>Lb11. plantarum 256 plasmid free growing, possibly contaminated<br>Lb12. reuteri DSM-20016 plasmid free growing, possibly contaminated<br>Lb23. reuteri 100-23-pSBLb4C15-red: no growth<br>Lb24. reuteri DSM-20016-pSBLb4C15-red no growth<br>Lb25. reuteri 100-23-pSBLbC2-red no growth<br>Lb28. reuteri 100-23-pVS2-red no growth<br><i>→ Do new competent cells and transformation immediately</i><br><br><b>Today:</b><br><b>Screening-pcr/Gel electrophoresis: </b><br>169. pSBLbEcC2-red (20 clones): No correct bands, just smudges.<br><i>→ Screen 20 new colonies for correct plasmid, use phusion polymerase and pEL-primers, giving some kind of (bigger) fragments regardless of whether the clone has the correct plasmid or not. </i><br></div>'; | ||
| + | } | ||
| + | else if(id == 'd2013731') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-07-31</h1><br><b>Name of participants:</b> Magnus, Pontus, Sabri, Thorsteinn, Nils, Malin, Alona Nyberg (AN), Christoffer Ahlström (CA), Anton Berglund (AB), Mikael Strandgren (MS), Jens Isaksson (JI), Marcus H, Karl H, Theodor L, Emil M, Victor S, Hampus.E<br><br><h2>Ongoing constructs</h2><br><b>E.coli:</b><br>19.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>20.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>108.pSBA15-J23106-B0034-IspA<br>109.pSBA15-K206000-B0034-IspA<br>110.pSBA15-K206001-B0034-IspA<br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pSB3T5-J23101-B0034-UGTCs2-6xHis<br>142.pSB3T5-CP1-B0034-UGTCs2-6xHis<br>143.pSB3T5-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSb1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>169. pSBLbEcC2-red<br>191. pSB1A2-fusionGFP<br>139. pSB3K3-J23101-B0032-BFP<br>192.pSB3K3-J23101-B0032-BFP<br>114. pSB1C3-B0034-TAL(MUT)<br>105. pSB3T5-J23101-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3K5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br>204. pEL3S17-J23110-HIS-STS<br>205.pSBLB4C15-CP1-B0034-HIS-TAL<br>206.pSB1C3-J23101-B0034-TAL<br><br><b>L. reuteri:</b><br>4.1 100-23 no plasmid<br>12.1 DSM 20016 no plasmid<br><br><h2>Todays work</h2><br><b>Screening:</b><br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pSB3T5-J23101-B0034-UGTCs2-6xHis<br>142.pSB3T5-CP1-B0034-UGTCs2-6xHis<br>176.pSB1C3-B0034-Exp-Linker-Miraculin-His<br>169. pSBLbEcC2-red (20 clones)<br> <br><b>Overnight:</b><br>142.pSB3T5-CP1-B0034-UGTCs2-6xHis<br>176.pSB1C3-B0034-Exp-Linker-Miraculin-His<br>Lb4.1 reuteri 100-23 no plasmid<br>Lb12.1 reuteri DSM 20016 no plasmid<br>172. pSB3T5-CP1-B0034-TAL (clone 3)<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS (clone 3 and 4)<br>189. pSB3K3-J23101-B0034-TAL (clone 2 and 3)<br><br><b>Ligation:</b><br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>36.pEL3S17-Red, 127. pSB1A3-B0034-UGTCs2-His, 92.BBa_J61002 (amp)-J23110 -><br>140.pEL3S17-Red-J23110-B0034-UGTCs2-6xHis <br>36.pEL3S17-Red, 127. pSB1A3-B0034-UGTCs2-His, 90.BBa_J61002 (amp)-J23101 -> 141.pEL3S17-Red-J23101-B0034-UGTCs2-6xHis<br><br><b>Transformation:</b><br>99. pCB1C3 - CrtO<br>190. pEL3K16 - CrtO<br>108.pSBA15-J23106-B0034-IspA<br>109.pSBA15-K206000-B0034-IspA<br>110.pSBA15-K206001-B0034-IspA<br>140.pEL3S17-Red-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-Red-J23101-B0034-UGTCs2-6xHis<br>143.pSB3T5-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>139. pSB3K3-J23101-B0032-BFP<br>192.pSB3K3-J23101<br><br><b>Re-Streak:</b><br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>191.1 pSB1A2-fusionGFP<br>191.2 pSB1A2-fusionGFP<br>191.3 pSB1A2-fusionGFP<br>191.4 pSB1A2-fusionGFP<br>139. pSB3K3-J23101-B0032-BFP<br>192.pSB3K3-J23101-B0032-BFP<br><br><b>Gel:</b><br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1 [Linear from PCR mutagenesis of 19] <br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1 [linear from PCR mutagenesis of 20]<br><br><b>Preparation for sequencing: </b><br>153. pSB1C3-j23110-meffBlue <br>155. pSB1C3-j23110-meffRed<br>156. pSB1C3-j23106-meffRed<br>157. pSB1C3-j23110-spisPink<br>159. pSB1C3-j23110-gfasPurple<br>160. pSB1C3-j23106-gfasPurple<br>150. pSB1C3-spispink<br>151. pSB1C3-meffblue<br>149. pSB1C3-meffred<br>148. pSB1C3-gfas purple<br><br><b>Assembly:</b><br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>204. pEL3S17-J23110-HIS-STS<br>205.pSBLB4C15-CP1-B0034-HIS-TAL<br>206.pSB1C3-J23101-B0034-TAL<br><br><b>PCR cloning and screening on gel of constructs:</b><br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br><br><h2>Results</h2><br><b>Screening:</b><br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis Failed<br>141.pSB3T5-J23101-B0034-UGTCs2-6xHis Failed<br>142.pSB3T5-CP1-B0034-UGTCs2-6xHis One colony succeded<br>Tranformations of 108, 109, 110, 177, 141, 180, and 182 failed.<br>Tranformations of 163, 178 and 179 succeeded.<br>Transformations of 181 and 140 remain inconclusive.<br><br><b>Screening of PCR products:</b><br>188.3 , 188.4, 189.2 and 189.3 have correct bands on the gel<br><br><h2>Other experiments</h2><br>LB-agar plates making: <br>Ampicillin, 28 plates<br>Chloramphenicol, 30 plates<br>Kanamycin, 31 plates<br>Spectinomycin, 26 plates<br>Tetracycline, 29 plates<br></div>'; | ||
| + | } | ||
| + | else if(id == 'd201381') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-08-01</h1><p>Name of participants: Marcus H, Karl H, Lovisa P, Emil M, Victor S, Hampus.E<br><br><h2>Ongoing constructs:</h2>' + | ||
| + | '114. pSB1C3-B0034-TAL(MUT)<br>105. pSB3T5-J23101-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3K5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>'+ | ||
| + | '137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>'+ | ||
| + | '187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br>204. pEL3S17-J23110-HIS-STS<br>205.pSBLB4C15-CP1-B0034-HIS-TAL<br>206.pSB1C3-J23101-B0034-TAL<br>'+ | ||
| + | |||
| + | '<h2>Todays work</h2><b>PCR screening and gel electrophorersis of construct:</b><br>185.(5-10) pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>'+ | ||
| + | |||
| + | '<h2>sequencing prep. of constructs:</h2><br>185.9-185.10<br>'+ | ||
| + | |||
| + | '<h2>Transformation of constructs:</h2><br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>204. pEL3S17-J23110-HIS-STS<br>205.pSBLB4C15-CP1-B0034-HIS-TAL<br>206.pSB1C3-J23101-B0034-TAL<br><br>'+ | ||
| + | |||
| + | '<h2>Results</h2>Construct 185, clone 9 and 10, bonds around 3000/4000 bp.<br><br>'+ | ||
| + | |||
| + | '<h2>Follow up</h2><br>- <br><br>'+ | ||
| + | |||
| + | '<h2>Other experiments</h2><br>-<br></p></div>'; | ||
| + | } | ||
| + | else if(id == 'd201382') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-08-02</h1><br><b>Name of participants: </b>Anton B, Christoffer A, Mikael S, Jens I, Marcus H, Karl H, Lovisa P, Emil M, Victor S, Hampus.E, Magnus, Pontus, Sabri, Nils<br><br><h2>Ongoing constructs:</h2><br><b>E.coli:</b><br>114. pSB1C3-B0034-TAL(MUT)<br>105. pSB3T5-J23101-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>108. pSBA15-J23106-B0034-IspA<br>109. pSBA15-K206000-B0034-IspA<br>110. pSBA15-K206001-B0034-IspA<br>115. pSB3K5-J23101-B0034-His-TAL<br>116. pSB3T5-J23110-B0034-His-TAL<br>119. pSB1C3-B0034-4CL<br>120. pSB1C3-B0034-HIS-4CL<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>140. pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141. pSB3T5-J23101-B0034-UGTCs2-6xHis<br>142. pSB3T5-CP1-B0034-UGTCs2-6xHis<br>163. pEL3S17-ZCD-His-B0034-UGTCs2-His<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>177. pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178. pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179. pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180. pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183. pEL3A15-B0034-CrtZ<br>184. pSB1C3-B0034-CrtZ<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br>191. pSB1A2-fusionGFP<br>193. pSB1C3-amilGFP<br>194. pSB1C3-B0034-amilCP<br>195. pSB1C3-cjBlue<br>196. pSB1C3-eforRed<br>197. pSB1C3-aeBlue<br>198. pSB1C3-B0034-amilGFP<br>199. pSb1C3-amajCFP<br>200. pSB1C3-B0034-asCP<br>202. pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>204. pEL3S17-J23110-HIS-STS<br>205. pSBLB4C15-CP1-B0034-HIS-TAL<br>206. pSB1C3-J23101-B0034-TAL<br>207. pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br><br><b>L. reuteri:</b><br>3. DSM 20016 pVS2<br>4. 100-23 no plasmid<br>12. DSM 20016 no plasmid<br>23. 100-23 pSBLb4C15-red<br>24. DSM 20016 pSBLb4C15-red<br>26. 100-23 pSBLbEcC2-red<br>27. DSM 20016 pSBLbEcC2-red<br>28. 100-23 pVS2<br><br><h2>Todays work</h2><br><b>Plasmid preparation: </b><br>139.1.1 pSB3K3-J23101-B0032-BFP <br>139.1.2 pSB3K3-J23101-B0032-BFP<br>191.1 pSB1A2-fusionGFP<br>191.2 pSB1A2-fusionGFP<br><br><b>Ligation:</b><br>4.pSB1A3-Red, 147.pSB1C3-B0034-Thioredoxin, 125.pSB1C3-ZCD-His-><br>208.pSB1A3-B0034-Thioredoxin-ZCD-His<br>36.pEL3S17-Red, 127. pSB1A3-B0034-UGTCs2-His, 90.BBa_J61002 (amp)-J23101 -> 141.pEL3S17-Red-J23101-B0034-UGTCs2-6xHis<br>152.pUC57-B0034-Exp-Linker-Miraculin-His(X,P), 36.pEL3S17-red, 103.pSB1K3-J23101 -> 177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br><br><b>Streaking of plasmid-free cells from frozen stock as control of C-resistance: </b><br>Lb4.1 reuteri. 100-23 no plasmid<br>Lb12.1 reuteri DSM 20016 no plasmid<br><br><b>Restreak of construct:</b><br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>204. pEL3S17-J23110-HIS-STS<br>205.pSBLB4C15-CP1-B0034-HIS-TAL<br>206.pSB1C3-J23101-B0034-TAL<br><br><b>Screening:</b><br>99. pCB1C3 - CrtO<br>190. pEL3K16 - CrtO<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br><br><b>Overnight:</b><br>99. pCB1C3 - CrtO<br>190. pEL3K16 - CrtO<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br><br><b>Gel electrophoresis:</b><br>PCR of 152. pUC57-B0034-Exp-Linker-Miraculin-Linker-His to extract Miraculin with biobrick prefix and suffix<br><br><h2>Results</h2><br><b>Today:</b><br><b>Plasmid preparation:</b> <br>139.1.1 pSB3K3-J23101-B0032-BFP: 51.9 ng/µl <br>139.1.2 pSB3K3-J23101-B0032-BFP: 53.6 ng/µl<br>191.1 pSB1A2-fusionGFP: 78.9 ng/µl<br>191.2 pSB1A2-fusionGFP: 74.1 ng/µl</div>' | ||
| + | } | ||
| + | else if(id == 'd201383') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-08-03</h1><br><b>Name of participants: </b>Lovisa P, Magnus, Pontus<br><br><h2>Other experiments</h2><br><b>Making competent cells (D5α)</b></div>' | ||
| + | } | ||
| + | else if(id == 'd201383') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-08-03</h1><br><b>Name of participants: </b>Lovisa P, Magnus, Pontus<br><br><h2>Other experiments</h2><br><b>Making competent cells (D5α)</b></div>' | ||
| + | } | ||
| + | else if(id == 'd201384') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Sunday 2013-08-04</h1><br><b>Name of participants: </b>Emil M, Magnus, Pontus<br><br><h2>Ongoing constructs:</h2><br>108.pSBA15-J23106-B0034-IspA<br>109.pSBA15-K206000-B0034-IspA<br>110.pSBA15-K206001-B0034-IspA<br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pSB3T5-J23101-B0034-UGTCs2-6xHis<br>142.pSB3T5-CP1-B0034-UGTCs2-6xHis<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSb1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br><br><h2>Todays work:</h2><br><b>Overnight:</b><br>10. D5α<br><br><h2>Characterization</h2><br><b>Detection of TAL activity through spectroscopy (day 1)</b><br><b>Contructs tested for TAL activity:</b><br>106.5 pSB3K3-J23110-B0034-TAL <br>106.6 pSB3K3-J23110-B0034-TAL<br>137.1 pSB3K3-CP8-B0034-TAL<br>189.3 pSB3K3-J23101-B0034-TAL<br><br><b>Negative controls:</b><br>43.7 pSB1C3-B0034-TAL<br>119.2 pSB1C3-B0034-4CL<br><br><b>Positive controls (p-coumaric acid added before dextraction):</b><br>43.7 pSB1C3-B0034-TAL<br>119.2 pSB1C3-B0034-4CL<br><br><h2Other experiments:</b><br><b>Competent cell test</b><br><br></div>' | ||
| + | } | ||
| + | else if(id == 'd201385') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-08-05</h1><br><b>Name of participants: </b>Anton B, Viktor T, Mikael S, Marcus H, Lovisa P, Emil M, Victor S, Hampus.E, Kristoffer L, Magnus, Pontus, Sabri, Malin, Niclas, Alexander, Thorsteinn<br><br><h2>Ongoing constructs:</h2><br><b>L. reuteri:</b><br>23. 100-23 pSBLb4C15-red<br>25. 100-23 pSBLbC2-red<br>28. 100-23 pVS2<br><br><b>E. coli:</b><br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSb1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br><br><h2>Todays work</h2><br><b>Competent cells</b><br>Lb4.1 reuteri 100-23 no plasmid<br><br><b>Lambda red transformation:</b><br>Crt<br><br><b>Transformation:</b><br>reuteri 100-23 pSB1C3-red (1 tube, to see if it would work in Lb, died in electroporation)<br>Lb23. reuteri 100-23 pSBLb4C15-red (5 tubes, 1 died in electroporation)<br>Lb25. reuteri 100-23 pSBLbC2-red (1 tube)<br>Lb28. reuteri 100-23 pVS2 (2 tubes, 1 died in electroporation)<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>184.pSB1C3-B0034-CrtZ<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br><br><b>Screening:</b><br>164.E.Coli Nissle-CrtY (clones from transformation plate)<br>201.E.Coli Nissle-CrtY-CrtI<br><br><h2>Characterization</h2><br><b>Detection of TAL activity through spectroscopy (day 2)</b><br><b>Contructs tested for TAL activity:</b><br>106.5 pSB3K3-J23110-B0034-TAL <br>106.6 pSB3K3-J23110-B0034-TAL<br>137.1 pSB3K3-CP8-B0034-TAL<br>189.3 pSB3K3-J23101-B0034-TAL<br><br><b>Negative controls:</b><br>43.7 pSB1C3-B0034-TAL<br>119.2 pSB1C3-B0034-4CL<br><br><b>Positive controls (p-coumaric acid added before dextraction):</b><br>43.7 pSB1C3-B0034-TAL<br>119.2 pSB1C3-B0034-4CL<br><br><h2>Other experiments</h2><br><b>Casting LB-agar plates: </b><br>Kanamycin, 63 plates<br>Chloramphenicol, 61 plates<br>Spectinomycin, 65 plates<br>Tetracycline, 32 plates<br>Ampicillin, 30 plates<br><br><b>Competent cells (D5α)</b></div>' | ||
| + | } | ||
| + | else if(id == 'd201386') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-08-06</h1><br><b>Name of participants:</b> Magnus, Pontus, Sabri, Malin, Niclas, Alexander, Thorsteinn, Marcus H, Lovisa P, Emil M, Victor S, Hampus.E, Kristoffer L, Anton B, Mikael S, Viktor T<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>57.3.pSB1C3-B0034-Idi<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSb1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>114. pSB1C3-B0034-TAL(MUT)<br>105. pSB3T5-J23101-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3K5-J23101-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br>204. pEL3S17-J23110-HIS-STS<br>205.pSBLB4C15-CP1-B0034-HIS-TAL<br>206.pSB1C3-J23110-B0034-TAL<br>224. pSB1C3-J23101-B0032-TAL<br>225. pEL3S17-J23101-B0032-TAL-B0034-4cl-B0034-STS<br>6. pSB1K3-red<br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>111. pSBLb4C15-red<br>139. pSB3K3-J23101-B0032-BFP<br>165. pSB1C3-RBS-tsPurle<br>166. pSB1C3-RBS-scOrange<br>167. pSB1C3-RBS-fwYellow<br>194. pSB1C3-B0034-amilCP<br>198. pSB1C3-B0034-amilGFP<br><br><b>L. reuteri:</b><br>23. 100-23 pSBLb4C15-red<br>25. 100-23 pSBLbC2-red<br>28. 100-23 pVS2 <br><br><h2>Todays work:</h2><br><b>Transformation:</b><br>109.pEL3S17-K206000-B0034-IspA<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin (E,P & E,X)<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>219.pSB1C3-Miraculin<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br><br><b>Restreak:</b><br>110.pEL3S17-K206001-B0034-IspA<br>115. pSB3K5-J23101-B0034-His-TAL<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>184.pSB1C3-B0034-CrtZ<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br><br><b>Digest:</b><br>6. pSB1K3-red<br>19.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>20.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>21.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>68.2 pSB3K3-CP1-B0032-BFP (EcoRI + PstI)<br>69.4 pSB3K3-CP8-B0032-BFP (EcoRI + PstI)<br>70.3 pSB3K3-CP11-B0032-BFP (EcoRI + PstI)<br>71.1 pSB3K3-CP29-B0032-BFP (EcoRI + PstI)<br>72.2 pSB3K3-CP30-B0032-BFP (EcoRI + PstI)<br>73.4 pSB3K3-CP41-B0032-BFP (EcoRI + PstI)<br>74. pSB3K3-CP44-B0032-BFP (EcoRI + PstI)<br>111. pSBLb4C15-red (XbaI + PstI)<br>165. pSB1C3-RBS-tsPurle (XbaI + PstI)<br>166. pSB1C3-RBS-scOrange (XbaI + PstI)<br>167. pSB1C3-RBS-fwYellow (XbaI + PstI)<br>194. pSB1C3-B0034-amilCP (XbaI + PstI)<br>198. pSB1C3-B0034-amilGFP (XbaI + PstI)<br><br><b>Ligation:</b><br>B0034-Thioredoxin, 1. pSB1C3-red (E, P) -> 147. pSB1C3-B0034-Thioredoxin<br>B0034-Thioredoxin, 1. pSB1C3-red (X, P) -> 147. pSB1C3-B0034-Thioredoxin<br><br><b>Gel: </b><br>164.E.Coli Nissle-CrtY (clones from transformation plate)<br>201.E.Coli Nissle-CrtY-CrtI<br><br><b>Screening:</b><br>164.E.Coli Nissle-CrtY (clones from transformation plate)<br>201.E.Coli Nissle-CrtY-CrtI<br>204.(1-4) pEL3S17-J23110-B0034-HIS-STS<br>171(1-4) pEL3S17-J23110-B0034-4CL-B0034-STS<br><br><b>Mutagenesis:</b><br>43.7 pSB1C3-B0034-TAL<br>57.3.pSB1C3-B0034-Idi -> 129.pSB1C3-Idi(mut)<br><br><v>Sequencing preparation:</b><br>139.1 pSB3K3-J23101-B0032-BFP<br>185.9 pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>185.10 pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br><br><b>O/N:</b><br>204.2 pEL3S17-J23110-B0034-HIS-STS<br>204.3 pEL3S17-J23110-B0034-HIS-STS<br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>119. pSB1C3-B0034-4CL (from 15)<br>111.7 pSBLb4C15-red<br><br><h2>Results:</h2><br><b>Gel:</b> <br>164.E.Coli Nissle-CrtY (clones from transformation plate) failed<br>201.E.Coli Nissle-CrtY-CrtI failed<br><br><b>Previous day:</b><br><b>Transformation: </b><br>No visual growth. Keeping them on growth for one more day.<br><br><h2>Characterization:</h2><br><b>Detection of TAL activity through spectroscopy (day 3)</b><br><b>Contructs tested for TAL activity:</b><br>106.5 pSB3K3-J23110-B0034-TAL <br>106.6 pSB3K3-J23110-B0034-TAL<br>137.1 pSB3K3-CP8-B0034-TAL<br>189.3 pSB3K3-J23101-B0034-TAL<br><br><b>Negative controls:</b><br>43.7 pSB1C3-B0034-TAL<br>119.2 pSB1C3-B0034-4CL<br><br><b>Positive controls (p-coumaric acid added before dextraction):</b><br>43.7 pSB1C3-B0034-TAL<br>119.2 pSB1C3-B0034-4CL<br><br><h2>Other Experiments:</h2><br>10.D5Alpha competence test<br><br><b>Preparation of buffers for Western Blot</b><br><br><b>Casting LB-agar plates:</b><br>Kanamycin, 33 plates<br>Spectinomycin, 31 plates<br><br></div>' | ||
| + | } | ||
| + | else if(id == 'd201387') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-08-07</h1><br><b>Name of participants: </b>- Viktor T, Anton B, Mikael S, Marcus H, Lovisa P, Emil M, Victor S, Hampus.E, Kristoffer L, Malin B, Magnus, Pontus, Sabri, Malin, Niclas, Alexander, Nils<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>6. pSB1K3-red<br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>92. J23110<br>111. pSBLb4C15-red<br>165. pSB1C3-RBS-tsPurple<br>166. pSB1C3-RBS-scOrange<br>167. pSB1C3-RBS-fwYellow<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>240. pSB1K3-J23110-RBS-tsPurple<br>241. pSB1K3-J23110-RBS-scOrange<br>242. pSB1K3-J23110-RBS-fwYellow<br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>114. pSB1C3-B0034-TAL(MUT)<br>105. pSB3T5-J23101-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3K5-J23101-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br>204. pEL3S17-J23110-HIS-STS<br>205.pSBLB4C15-CP1-B0034-HIS-TAL<br>206.pSB1C3-J23110-B0034-TAL<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut)<br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>148. pSB1C3-RBS-gfasPurple<br>150. pSB1C3-RBS-spisPink<br>151. pSB1C3-RBS-meffBlue<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSb1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>224. pSB1C3-J23101-B0032-TAL<br>225. pEL3S17-J23101-B0032-TAL-B0034-4cl-B0034-STS<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>243. pSB1A2-LacI<br>244. pSB1C3-Promoter-RBS-Bleomysin<br>245. pSB1C3-J23110-B0034-amajCP(amajLime)<br>246. pSB1C3-J23110-b0034-gfasPurple<br><br><h2>Todays work</h2><br><b>Assembly:</b><br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-meffRed à 155. pSB1C3-j23110-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-meffRed à 156. pSB1C3-j23106-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-spisPink à 157. pSB1C3-j23110-RBS-spisPink<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-spisPink à 158. pSB1C3-j23106-RBS-spisPink<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>240. pSB1K3-J23110-RBS-tsPurple*<br>241. pSB1K3-J23110-RBS-scOrange*<br>242. pSB1K3-J23110-RBS-fwYellow*<br><i>*Will eventually be tried in shuttle vector, but that has C-resistance, so a subcloning to K is necessary first, or a modification to change the resistance of the shuttle vector.</i><br><br><b>Transformation:</b><br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>240. pSB1K3-J23110-RBS-tsPurple<br>241. pSB1K3-J23110-RBS-scOrange<br>242. pSB1K3-J23110-RBS-fwYellow<br>108.pEL3S17-J23106-B0034-IspA<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin (E,P & E,X)<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>243. pSB1A2-LacI<br>244. pSB1C3-Promoter-RBS-Bleomysin<br>148. pSB1C3-RBS-gfasPurple<br>150. pSB1C3-RBS-spisPink<br>151. pSB1C3-RBS-meffBlue<br><br><b>Spreading: </b><br>60. PK401 dam- no plasmid<br>111.7 pSBLb4C15-red<br><br><b>Restreak:</b><br>109.pEL3S17-K206000-B0034-IspA<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>219.pSB1C3-Miraculin<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>228.pEL3A17-Blue<br>229.pEL3K17-Blue<br>230.pEL3S15-Blue<br>231.pEL3C17-Blue<br><br><b>O/N & Screening:</b><br>110.pEL3S17-K206001-B0034-IspA<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pBAD/AraC-RBS-Luciferas (2012 years kit)<br>227.pBAD/AraC-RBS-Luciferas (2013 years kit)<br>245. pSB1C3-J23110-B0034-amajCP(amajLime)<br>246. pSB1C3-J23110-b0034-gfasPurple<br><br><b>Gel:</b><br>164.E.Coli Nissle-CrtY (clones from transformation plate)<br>201.E.Coli Nissle-CrtY-CrtI<br>129.pSB1C3-Idi(mut)<br><br><h2>Results</h2><br><b>Transformation (Monday):</b><br>Still no visual growth. <br><br><b>SDS-page failed (probably due to faulty buffers, investigate the running buffer first)</b><br><br><h2>Characterization experiments</h2><br><b>SDS-page on constructs:</b><br>43.7 pSB1C3-B0034-TAL<br><br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>119. pSB1C3-B0034-4CL (from 15)<br><br><h2>Other experiments</h2><br><b>Sequencing preparation:</b><br>69. pSB3K3-CP8-B0032-BFP*<br><i>*since the existing sequencing data doesn’t cover the whole insert with sufficient accuracy. There may be a mutation at the end of BFP, but that is more than 800bp after the single primer. Was not sent today, not sequencing tuesday.</i></div>' | ||
| + | } | ||
| + | else if(id == 'd201388') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-08-08</h1><br><b>Name of participants:</b> Magnus, Pontus, Sabri, Malin, Niclas, Alexander, Nils, Marcus H, Lovisa P, Emil M, Victor S, Hampus.E, Kristoffer L, Malin B, Stephanie H, Mikael S, Anton B<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>60. Strain PK401 dam- no plasmid<br>111.7 pSBLb4C15-red<br>139. pSB3K3-J23101.B0032-BFP<br>91. BBa_J61002-J23106<br>92. BBa_J61002-J23110<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut)<br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>148. pSB1C3-RBS-gfasPurple<br>150. pSB1C3-RBS-spisPink<br>151. pSB1C3-RBS-meffBlue<br>165. pSB1C3-RBS-tsPurple<br>166. pSB1C3-RBS-scOrange<br>167. pSB1C3-RBS-fwYellow<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSb1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>243. pSB1A2-LacI<br>244. pSB1C3-Promoter-RBS-Bleomysin<br>245. pSB1C3-J23110-B0034-amajCP(amajLime)<br>246. pSB1C3-J23110-b0034-gfasPurple<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow<br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>114. pSB1C3-B0034-TAL(MUT)<br>105. pSB3T5-J23101-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3K5-J23101-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br>204. pEL3S17-J23110-HIS-STS<br>205.pSBLB4C15-CP1-B0034-HIS-TAL<br>206.pSB1C3-J23110-B0034-TAL<br>224. pSB1C3-J23101-B0032-TAL<br>225. pEL3S17-J23101-B0032-TAL-B0034-4cl-B0034-STS<br><br><h2>Todays work:</h2><br><b>Transformation:</b><br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin (E,P & E,X)<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>no nr. pSB1c3-Thioredoxin<br>no nr. pEL3S17-Thioredoxin<br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-meffRed à 155. pSB1C3-j23110-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-meffRed à 156. pSB1C3-j23106-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-spisPink à 157. pSB1C3-j23110-RBS-spisPink<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-spisPink à 158. pSB1C3-j23106-RBS-spisPink<br><br><b>Re-streak:</b><br>108.pEL3S17-J23106-B0034-IspA<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1 <br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>243. pSB1A2-LacI<br>244. pSB1C3-Promoter-RBS-Bleomysin<br>148. pSB1C3-RBS-gfasPurple<br>150. pSB1C3-RBS-spisPink<br>151. pSB1C3-RBS-meffBlue<br>228.pEL3A17-Blue<br>229.pEL3K17-Blue<br>230.pEL3S15-Blue<br>231.pEL3C17-Blue<br><br><bPlasmid prepp</b><br>226. pBAD/AraC-RBS-Luciferas (2012 years kit)<br>227.pBAD/AraC-RBS-Luciferas (2013 years kit)<br>245. pSB1C3-J23110-B0034-amajCP(amajLime)<br>246. pSB1C3-J23110-b0034-gfasPurple<br><br><b>O/N screening:</b><br>109.pEL3S17-K206000-B0034-IspA<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Link<br>er-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>219.pSB1C3-Miraculin<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br><br><b>Two step Assembly</b><br>165.1 pSB1C3-RBS-tsPurple, 91. BBa_J61002-J23106 à 247. pSB1C3-J23110-RBS-tsPurple<br>165.1 pSB1C3-RBS-tsPurple, 92. BBa_J61002-J23110 à 248. pSB1C3-J23106-RBS-tsPurple<br>166.1 pSB1C3-RBS-scOrange, 91. BBa_J61002-J23106 à 249. pSB1C3-J23110-RBS-scOrange<br>166.1 pSB1C3-RBS-scOrange, 92. BBa_J61002-J23110 à 250. pSB1C3-J23106-RBS-scOrange<br>167.1 pSB1C3-RBS-fwYellow, 91. BBa_J61002-J23106 à 251. pSB1C3-J23110-RBS-fwYellow<br>167.1 pSB1C3-RBS-fwYellow, 92. BBa_J61002-J23110 à 252. pSB1C3-J23106-RBS-fwYellow<br><br><b>Overnight (screening)</b><br>1. pSB1C3-red<br><br><b>O/N</b><br>60. Strain PK401 dam- no plasmid<br>111.7 pSBLb4C15-red<br><br><b>Streak from frozen stock</b><br>3. pSB3T5-red<br>6. pSB1K3-red<br>42. pSB4S15-red<br>77. pSB4C15-red<br><br><h2>Results:</h2><br><b>One of the SDS-gels failed, resulting in no good gel to stain (faulty buffers, continue to search for sources of errors)</b><br><br><b>The succesfull gel was prepared with control buffers.</b><br><br><b>Transformation of promotors with BFP in the shuttle vector, for characterization in Lactobacillus: Some growth, only white colonies, will let them grow overnight again. </b><br><br><h2>Characterization experiments:</h2><br><b>Westernblot </b><br><b>SDS-page on constructs:</b><br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>119. pSB1C3-B0034-4CL (from 15)<br><br><b>Electro transfer over night</b><br><br><b>Staining of control gel</b><br><br><b>Detection of TAL activity through spectroscopy (day 4)</b></div>' | ||
| + | } | ||
| + | else if(id == 'd201389') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-08-09</h1><br><b>Name of participants:</b> Stephanie H, Mikael S, Anton B, Anders E, Magnus, Pontus, Sabri, Malin, Alexander, Nils, Christoffer<br><br><h2>Ongoing constructs:</h2><br>60. PK401 dam- no plasmid<br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>139. pSB3K3-J23101-B0034-BFP<br>111.7 pSBLb4C15-red<br>91. BBa_J61002-J23106<br>92. BBa_J61002-J23110 <br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut)<br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>148. pSB1C3-RBS-gfasPurple<br>150. pSB1C3-RBS-spisPink<br>151. pSB1C3-RBS-meffBlue<br>165. pSB1C3-RBS-tsPurple<br>166. pSB1C3-RBS-scOrange<br>167. pSB1C3-RBS-fwYellow<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>244. pSB1C3-Promoter-RBS-Bleomysin<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow<br><br><h2>Todays work</h2><br><b>Streak from frozen stock:</b><br>60. PK401 dam- no plasmid<br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>139. pSB3K3-J23101-B0034-BFP<br><br><b>O/N:</b><br>60.3 PK401 dam- no plasmid*<br><i>*The O/N made on thursday used MRS, so it had to be remade from restreaks.</i><br><br><b>Plasmid preparation:</b><br>111.7 pSBLb4C15-red<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br><br><b>Restreak:</b><br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin (E,P & E,X)<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>no nr. pEL3S17-Thioredoxin<br><br><b>Amplification PCR</b><br>134. B0034-CBP<br>135. B0034-CSP<br>254. Gus<br>257. p-pem<br>258. pem<br><br><b>Mutagenesis</b><br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br><br><b>O/N Screening:</b><br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Link<br>er-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>219.pSB1C3-Miraculin<br>221. pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>201. E.Coli. Nissle - CrtY-CrtI<br>243. pSB1A2-LacI<br>244. pSB1C3-Promoter-RBS-Bleomysin<br>148. pSB1C3-RBS-gfasPurple<br>150. pSB1C3-RBS-spisPink<br>151. pSB1C3-RBS-meffBlue<br>228.pEL3A17-Blue<br>229.pEL3K17-Blue<br>230.pEL3S15-Blue<br>231.pEL3C17-Blue<br><br><b>Send to Linköping, as plasmid prepp and agar stabb</b><br>226.pSB1C3-pBAD/AraC-RBS-Luciferas (2012 years kit)<br>227.pSB1C3-pBAD/AraC-RBS-Luciferas (2013 years kit)<br><br><b>Transformation</b><br>165.1 pSB1C3-RBS-tsPurple, 91. BBa_J61002-J23106 à 247. pSB1C3-J23110-RBS-tsPurple<br>165.1 pSB1C3-RBS-tsPurple, 92. BBa_J61002-J23110 à 248. pSB1C3-J23106-RBS-tsPurple<br>166.1 pSB1C3-RBS-scOrange, 91. BBa_J61002-J23106 à 249. pSB1C3-J23110-RBS-scOrange<br>166.1 pSB1C3-RBS-scOrange, 92. BBa_J61002-J23110 à 250. pSB1C3-J23106-RBS-scOrange<br>167.1 pSB1C3-RBS-fwYellow, 91. BBa_J61002-J23106 à 251. pSB1C3-J23110-RBS-fwYellow<br>167.1 pSB1C3-RBS-fwYellow, 92. BBa_J61002-J23110 à 252. pSB1C3-J23106-RBS-fwYellow<br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-meffRed à 155. pSB1C3-j23110-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-meffRed à 156. pSB1C3-j23106-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-spisPink à 157. pSB1C3-j23110-RBS-spisPink<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-spisPink à 158. pSB1C3-j23106-RBS-spisPink<br><br><br><h2>Results</h2><br><b>Plasmid preparation:</b><br>111.7 pSBLb4C15-red: 30,6 ng/µl<br><br><br><b>O/N:</b><br>221 failed, redo.<br><br><b>O/N Screening:</b><br>108.pEL3S17-J23106-B0034-IspA - Screening gel fail<br>109.pEL3S17-K206000-B0034-IspA - Screening gel fail<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis - <br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis - <br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His - Screening gel fail<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His - <br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1 - Screening gel fail<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Link<br>er-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1 - Screening gel fail<br>219.pSB1C3-Miraculin - <br>221. pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>201. E.Coli. Nissle - CrtY-CrtI - Screening gel fail</div>' | ||
| + | } | ||
| + | else if(id == 'd2013810') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-08-10</h1><br><b>Name of participants: </b>Anders Edlund, Marcus H, Hampus.E, Kristoffer L, Ken B-A, Nils, Alexander, Sabri, Pontus<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>134. B0034-CBP<br>135. B0034-CSP<br>254. Gus<br>257. p-Toxin-Antitoxin<br>258. Toxin-Antitoxin<br>114. pSB1C3-B0034-TAL(MUT)<br>105. pSB3T5-J23101-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3K5-J23101-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br>204. pEL3S17-J23110-HIS-STS<br>205.pSBLB4C15-CP1-B0034-HIS-TAL<br>206.pSB1C3-J23110-B0034-TAL<br>224. pSB1C3-J23101-B0032-TAL<br>225. pEL3S17-J23101-B0032-TAL-B0034-4cl-B0034-STS<br>259. pEL3S17-B0034-4CL-B0034-STS<br>114. pSB1C3-B0034-TAL(MUT)<br>105. pSB3T5-J23101-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3K5-J23101-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br>204. pEL3S17-J23110-HIS-STS<br>205.pSBLB4C15-CP1-B0034-HIS-TAL<br>206.pSB1C3-J23110-B0034-TAL<br>224. pSB1C3-J23101-B0032-TAL<br>225. pEL3S17-J23101-B0032-TAL-B0034-4cl-B0034-STS<br>259. pEL3S17-B0034-4CL-B0034-STS<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut)<br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br><br><b>E. coli PK401 dam-:</b><br>60. no plasmid<br><br><h2>Todays work:</h2><br><b>Gel-electrophoresis</b><br>134. B0034-CBP*<br>135. B0034-CSP*<br>254. Gus<br>257. p-Toxin-Antitoxin<br>258. Toxin-Antitoxin<br>*PCR 99% failed<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>115. pSB3K5-J23101-B0034-His-TAL<br>114. pSB1C3-B0034-TAL(MUT)<br>120.pSB1C3-B0034-HIS-4CL<br><br><b>Amplification PCR</b><br>134. B0034-CBP<br>135. B0034-CSP<br><br><b>ON:</b><br>60. E. coli dam-<br><br><b>Assembly and subcloning:</b><br>205.pSBLB4C15-CP1-B0034-HIS-TAL<br>206.pSB1C3-J23110-B0034-TAL<br>225. pEL3S17-J23101-B0032-TAL-B0034-4cl-B0034-STS<br>259. pEL3S17-B0034-4CL-B0034-STS<br>127 + 103 + 36 -> 253. pEL3S17-J23101-B0034-UGTCs2-His<br>152 + 61 + 111 -> 181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>152 + 62 + 111 -> 182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>95 + 58 + 36 -> 110.pEL3S17-K206001-B0034-IspA<br>CrtZ + 1 -> 184.pSB1C3-B0034-CrtZ<br>Miraculin + 1 -> 219.pSB1C3-Miraculin<br><br><b>O/N Screening:</b><br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Link<br>er-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>219.pSB1C3-Miraculin<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>201. E.Coli. Nissle - CrtY-CrtI<br><br><b>DPN1</b><br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br><br><h2>Results:</h2><br><b>Gel electrophoresis:</b><br>115 good, to sequencing<br>114 good, to sequencing<br>120 pick new clones and screen<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268 - Succes<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1 - fail<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1 -fail<br><br><b>Characterization experiments</b><br><b>Detection of TAL activity through spectroscopy (day 5)</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013811') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Sunday 2013-08-11</h1><br><b>Name of participants: </b>Anders E, Mikael S, Nafisa B, Nils, Alexander, Sabri, Malin<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>60. PK401 dam- no plasmid<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>91. BBa_J61002-J23106<br>92. BBa_J61002-J23110 <br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>148. pSB1C3-RBS-gfasPurple<br>150. pSB1C3-RBS-spisPink<br>151. pSB1C3-RBS-meffBlue<br>165. pSB1C3-RBS-tsPurple<br>166. pSB1C3-RBS-scOrange<br>167. pSB1C3-RBS-fwYellow<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>244. pSB1C3-Promoter-RBS-Bleomysin<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br><br><b>E. coli PK401 dam-:</b><br>111.7 pSBLb4C15-red<br>181. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br><br><h2>Todays work:</h2><br><b>Competent cells:</b><br>60. PK401 dam- noplasmid<br><br><b>Transformation:</b><br>275. PK401 dam- pSBLb4C15-red (dam-)<br>276. PK401 dam- pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His (dam-)<br>277. PK401 dam- pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His (dam-)<br><br><b>Gel electrophoresis:</b><br>111.7 pSBLb4C15 cut with E and P<br>and more.<br><br><b>Digest:</b><br>111.7 pSBLb4C15 cut with E and P<br>111.7 pSBLb4C15 cut with SacI and SalI<br>254. Gus P+X <br>257. p-Toxin-Antitoxin P+X <br>258. Toxin-Antitoxin P+X<br>134. B0034-CBP E+S<br>1. pSB1C3-red E+S<br>3. pSB3T5-red E+P<br><br><b>PCR purification:</b><br>254. pSB1C3-Gus<br>257. pSB1C3-p-Toxin-Antitoxin (with promotor)<br>258. pSB1C3-Toxin-Antitoxin<br>134. pSB1C3-B0034-CBP<br><br><b>PCR:</b><br>Lb2. reuteri pLUL631 (erythromycin resistance cassette, with primers pLUL-E-sc_F and pLUL-E-sl_R)<br><br><b>Plasmidprepp:</b><br>108.2.pEL3S17-J23106-B0034-IspA<br>108.3.pEL3S17-J23106-B0034-IspA<br>148. pSB1C3-RBS-gfasPurple<br>150. pSB1C3-RBS-spisPink<br>151. pSB1C3-RBS-meffBlue<br>163.1.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>243. pSB1A2-LacI<br>244. pSB1C3-Promoter-RBS-Bleomysin<br>228.pEL3A17-Blue<br>229.pEL3K17-Blue<br>230.pEL3S15-Blue<br>231.pEL3C17-Blue<br><br><b>Two part assembly:</b><br>165.1 pSB1C3-RBS-tsPurple, 91. BBa_J61002-J23106 à 247. pSB1C3-J23110-RBS-tsPurple<br>165.1 pSB1C3-RBS-tsPurple, 92. BBa_J61002-J23110 à 248. pSB1C3-J23106-RBS-tsPurple<br>166.1 pSB1C3-RBS-scOrange, 91. BBa_J61002-J23106 à 249. pSB1C3-J23110-RBS-scOrange<br>166.1 pSB1C3-RBS-scOrange, 92. BBa_J61002-J23110 à 250. pSB1C3-J23106-RBS-scOrange<br>167.1 pSB1C3-RBS-fwYellow, 91. BBa_J61002-J23106 à 251. pSB1C3-J23110-RBS-fwYellow<br>167.1 pSB1C3-RBS-fwYellow, 92. BBa_J61002-J23110 à 252. pSB1C3-J23106-RBS-fwYellow<br><br><b>O/N:</b><br>222.1.pSB3K3-CP1-B0034-UGTCs2-His<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His<br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis<br><br><b>Sekvensprepp:</b><br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br><br><b>Re-circularization:</b><br>202.pSB4C5.CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268 <br><br><b>Re-streak</b><br>165.1 pSB1C3-RBS-tsPurple, 91. BBa_J61002-J23106 à 247. pSB1C3-J23110-RBS-tsPurple<br>165.1 pSB1C3-RBS-tsPurple, 92. BBa_J61002-J23110 à 248. pSB1C3-J23106-RBS-tsPurple<br>166.1 pSB1C3-RBS-scOrange, 91. BBa_J61002-J23106 à 249. pSB1C3-J23110-RBS-scOrange<br>166.1 pSB1C3-RBS-scOrange, 92. BBa_J61002-J23110 à 250. pSB1C3-J23106-RBS-scOrange<br>167.1 pSB1C3-RBS-fwYellow, 91. BBa_J61002-J23106 à 251. pSB1C3-J23110-RBS-fwYellow<br>167.1 pSB1C3-RBS-fwYellow, 92. BBa_J61002-J23110 à 252. pSB1C3-J23106-RBS-fwYellow<br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-meffRed à 155. pSB1C3-j23110-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-meffRed à 156. pSB1C3-j23106-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-spisPink à 157. pSB1C3-j23110-RBS-spisPink<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-spisPink à 158. pSB1C3-j23106-RBS-spisPink<br><br><h2>Results:</h2><br><b>Gel electrophoresis:</b><br>111.7 pSBLb4C15 cut with E and P: correct bands!<br>and more.</div>' | ||
| + | } | ||
| + | else if(id == 'd2013812') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-08-12</h1><br><b>Name of participants: </b>Alexander, Sabri, Malin, Pontus, Niclas, Sabri, Hampus.E, Theodor L, Karl H, Ken B, Kristoffer L, Victor S, Marcus H, Lovisa P, Anders E, Mikael S, Nafisa B, Viktor B, Viktor T, Christoffer A, Stephanie H, Anton B<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>149. pSB1C3-B0034-Thioredoxin<br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>202. pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>114. pSB1C3-B0034-TAL-mut<br>115. pSB3K3-J23101-B0034-His-TAL<br>204. pEL3S17-J23110-B0034-HIS-STS<br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br><br><b>E. coli PK401 dam-:</b><br>111.7 pSBLb4C15-red<br>181. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br><br><h2>Todays work</h2><br><b>Ligation:</b><br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>208.pSB1A3-B0034-Thioredoxin-ZCD-His<br>272.pSB1K3-B0034-Thioredoxin-ZCD-His<br>273.pSB3T5-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>134. pSB1C3-B0034-CBP<br>254. pSB1C3-Gus<br>257. pSB1C3-p-Toxin-Antitoxin (with promotor)<br>258. pSB1C3-Toxin-Antitoxin<br>233. pSBLb4C15-CP1-B0034-BFP<br>234. pSBLb4C15-CP8-B0034-BFP<br>235. pSBLb4C15-CP11-B0034-BFP<br>236. pSBLb4C15-CP29-B0034-BFP<br>237. pSBLb4C15-CP30-B0034-BFP<br>238. pSBLb4C15-CP41-B0034-BFP<br>239. pSBLb4C15-CP44-B0034-BFP<br><br><b>Digest:</b><br>4.pSB1A3-red<br>6.pSB1K3<br>125. pSB1C3-ZCD-His<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br><br><b>Re-streak:</b><br>110.pEL3S17-K206001-B0034-IspA<br>219.pSB1C3-Miraculin<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>275 PK401 dam- pSBLb4C15-red (dam-)<br>276. PK401 dam- pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His (dam-)<br>277. PK401 dam- pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His (dam-)<br><br><b>O/N Screening:</b><br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br>222.1.pSB3K3-CP1-B0034-UGTCs2-His<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His<br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>225. pEL3S17 -J23101-B0034-TAL-B0034-4CL-B0034-STS<br><br><b>Gel electrophoresis:</b><br>Lb2. reuteri pLUL631 (erythromycin resistance cassette, with primers pLUL-E-sc_F and pLUL-E-sl_R)<br>CBP<br>Mcherry<br>Pem<br>Pemp<br><br><b>Transformation</b><br>134. pSB1C3-CBP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>233. pSBLb4C15-CP1-B0034-BFP<br>234. pSBLb4C15-CP8-B0034-BFP<br>235. pSBLb4C15-CP11-B0034-BFP<br>236. pSBLb4C15-CP29-B0034-BFP<br>237. pSBLb4C15-CP30-B0034-BFP<br>238. pSBLb4C15-CP41-B0034-BFP<br>239. pSBLb4C15-CP44-B0034-BFP<br><br><b>Spreading:</b><br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br><br><b>Plasmidprepp:</b><br>222.1.pSB3K3-CP1-B0034-UGTCs2-His<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His<br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>129. pSB1C3-B0034-Idi (mut)<br>149. pSB1C3-B0034-Thioredoxin<br><br><b>Sequence prepp:</b><br>129. pSB1C3-B0034-Idi (mut)<br>149. pSB1C3-B0034-Thioredoxin<br><br><b>Transformation:</b><br>202. pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br><br><b>PCR:</b><br>Lb2. reuteri pLUL631 (erythromycin resistance cassette, with primers pLUL-E-sc_F and pLUL-E-sl_R)<br>CBP<br>Mcherry<br>Pem<br>Pemp<br><br><h2>Results:</h2><br>222.1.pSB3K3-CP1-B0034-UGTCs2-His failed<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His failed<br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis failed<br><br><b>Gel electrophoresis:</b><br>Lb2. reuteri pLUL631 (erythromycin resistance cassette, with primers pLUL-E-sc_F and pLUL-E-sl_R)<br><br><h2>Characterization</h2><br><b>Characterization of Resveratrol production through spectroscopy and HPLC</b><br><br><h2>Other experiments</h2><br><b>Mixing new buffers for western blot</b><br><br><b>Casting LB-agar plates:</b><br>Chloramphenicol: 55</div>' | ||
| + | } | ||
| + | else if(id == 'd2013812') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-08-12</h1><br><b>Name of participants: </b>Alexander, Sabri, Malin, Pontus, Niclas, Sabri, Hampus.E, Theodor L, Karl H, Ken B, Kristoffer L, Victor S, Marcus H, Lovisa P, Anders E, Mikael S, Nafisa B, Viktor B, Viktor T, Christoffer A, Stephanie H, Anton B<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>149. pSB1C3-B0034-Thioredoxin<br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>202. pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>114. pSB1C3-B0034-TAL-mut<br>115. pSB3K3-J23101-B0034-His-TAL<br>204. pEL3S17-J23110-B0034-HIS-STS<br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br><br><b>E. coli PK401 dam-:</b><br>111.7 pSBLb4C15-red<br>181. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br><br><h2>Todays work</h2><br><b>Ligation:</b><br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>208.pSB1A3-B0034-Thioredoxin-ZCD-His<br>272.pSB1K3-B0034-Thioredoxin-ZCD-His<br>273.pSB3T5-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>134. pSB1C3-B0034-CBP<br>254. pSB1C3-Gus<br>257. pSB1C3-p-Toxin-Antitoxin (with promotor)<br>258. pSB1C3-Toxin-Antitoxin<br>233. pSBLb4C15-CP1-B0034-BFP<br>234. pSBLb4C15-CP8-B0034-BFP<br>235. pSBLb4C15-CP11-B0034-BFP<br>236. pSBLb4C15-CP29-B0034-BFP<br>237. pSBLb4C15-CP30-B0034-BFP<br>238. pSBLb4C15-CP41-B0034-BFP<br>239. pSBLb4C15-CP44-B0034-BFP<br><br><b>Digest:</b><br>4.pSB1A3-red<br>6.pSB1K3<br>125. pSB1C3-ZCD-His<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br><br><b>Re-streak:</b><br>110.pEL3S17-K206001-B0034-IspA<br>219.pSB1C3-Miraculin<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>275 PK401 dam- pSBLb4C15-red (dam-)<br>276. PK401 dam- pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His (dam-)<br>277. PK401 dam- pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His (dam-)<br><br><b>O/N Screening:</b><br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br>222.1.pSB3K3-CP1-B0034-UGTCs2-His<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His<br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>225. pEL3S17 -J23101-B0034-TAL-B0034-4CL-B0034-STS<br><br><b>Gel electrophoresis:</b><br>Lb2. reuteri pLUL631 (erythromycin resistance cassette, with primers pLUL-E-sc_F and pLUL-E-sl_R)<br>CBP<br>Mcherry<br>Pem<br>Pemp<br><br><b>Transformation</b><br>134. pSB1C3-CBP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>233. pSBLb4C15-CP1-B0034-BFP<br>234. pSBLb4C15-CP8-B0034-BFP<br>235. pSBLb4C15-CP11-B0034-BFP<br>236. pSBLb4C15-CP29-B0034-BFP<br>237. pSBLb4C15-CP30-B0034-BFP<br>238. pSBLb4C15-CP41-B0034-BFP<br>239. pSBLb4C15-CP44-B0034-BFP<br><br><b>Spreading:</b><br>68. pSB3K3-CP1-B0032-BFP<br>69. pSB3K3-CP8-B0032-BFP<br>70. pSB3K3-CP11-B0032-BFP<br>71. pSB3K3-CP29-B0032-BFP<br>72. pSB3K3-CP30-B0032-BFP<br>73. pSB3K3-CP41-B0032-BFP<br>74. pSB3K3-CP44-B0032-BFP<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br><br><b>Plasmidprepp:</b><br>222.1.pSB3K3-CP1-B0034-UGTCs2-His<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His<br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>129. pSB1C3-B0034-Idi (mut)<br>149. pSB1C3-B0034-Thioredoxin<br><br><b>Sequence prepp:</b><br>129. pSB1C3-B0034-Idi (mut)<br>149. pSB1C3-B0034-Thioredoxin<br><br><b>Transformation:</b><br>202. pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br><br><b>PCR:</b><br>Lb2. reuteri pLUL631 (erythromycin resistance cassette, with primers pLUL-E-sc_F and pLUL-E-sl_R)<br>CBP<br>Mcherry<br>Pem<br>Pemp<br><br><h2>Results:</h2><br>222.1.pSB3K3-CP1-B0034-UGTCs2-His failed<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His failed<br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis failed<br><br><b>Gel electrophoresis:</b><br>Lb2. reuteri pLUL631 (erythromycin resistance cassette, with primers pLUL-E-sc_F and pLUL-E-sl_R)<br><br><h2>Characterization</h2><br><b>Characterization of Resveratrol production through spectroscopy and HPLC</b><br><br><h2>Other experiments</h2><br><b>Mixing new buffers for western blot</b><br><br><b>Casting LB-agar plates:</b><br>Chloramphenicol: 55</div>' | ||
| + | } | ||
| + | else if(id == 'd2013813') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-08-13</h1><br><b>Name of participants: </b>Anders Edlund, Mikael Strandgren, Anton Berglund, Viktor Törnblom, Viktor Blomkvist, Christoffer Ahlström, Stephanie Herman, Hampus.E, Karl H, Ken B, Kristoffer L, Victor S, Marcus H, Theodor L. Lovisa P, Alexander, Malin, Pontus, Niclas, Christoffer, Nils, Sabri<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br><br><b>E. coli PK401 dam-:</b><br>111.7 pSBLb4C15-red<br>181. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>114.(2-3) pSB1C3-B0034-TAL-mut<br>115.(1-3) pSB3K3-J23101-B0034-His-TAL<br>204.(1-3) pEL3S17-J23110-B0034-HIS-STS<br><br><h2>Todays work:</h2><br><b>O/N Screening glycerolstock:</b><br>110.pEL3S17-K206001-B0034-IspA<br>219.pSB1C3-Miraculin<br>253.pEL3S17-J23101-B0034-UGTCs2-His<br><br><b>Assembly</b><br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-meffRed à 155. pSB1C3-j23110-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-meffRed à 156. pSB1C3-j23106-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-spisPink à 157. pSB1C3-j23110-RBS-spisPink<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-spisPink à 158. pSB1C3-j23106-RBS-spisPink<br><br><b>Ligation:</b><br>134.1 pSB1C3-CBP (134.1 from digested plasmids [PCR product])<br>134.2 pSB1C3-CBP (134.2 from digested plasmids [PCR product])<br>254.1 pSB1C3-gus (254.1 from digested plasmids [PCR product])<br>254.1 pSB1C3-gus (254.2 from digested plasmids [PCR product])<br>255.1 pSBLb4C15-pemp (257.1 from digest plasmids [PCR product])<br>255.2 pSBLb4C15-pemp (257.2 from digest plasmids [PCR product])<br>256.1 pSBLb4C15-pem (258.1 from digested plasmids [PCR product])<br>256.2 pSBLb4C15-pem (258.2 from digested plasmids [PCR product])<br><br><b>Plasmidprepp:</b><br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>222.1.pSB3K3-CP1-B0034-UGTCs2-His<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His<br><br><b>Sequencing prep:</b><br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>114.(2-3) pSB1C3-B0034-TAL-mut<br>115.(1-3) pSB3K3-J23101-B0034-His-TAL<br>204.(1-3) pEL3S17-J23110-B0034-HIS-STS<br>108.2.pEL3S17-J23106-B0034-IspA<br>108.3.pEL3S17-J23106-B0034-IspA<br>163.1.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>222.1.pSB3K3-CP1-B0034-UGTCs2-His<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His<br><br><b>Re-streaking:</b><br>134. pSB1C3-CBP (CBP1 [134.1] from digested plasmids [PCR-product])<br>134. pSB1C3-CBP (CBP2 [134.2] from digested plasmids [PCR-product])<br>254. pSB1C3-gus (gus1 [254.1] from digested plasmids [PCR-product])<br>254. pSB1C3-gus (gus2 [254.2] from digested plasmids [PCR-product])<br>254. pSB1C3-gus (gus3 [254.3] from digested plasmids [PCR-product])<br>255. pSBLb4C15-pemp (pemp1 [257.1] from digested plasmids [PCR-product])<br>255. pSBLb4C15-pemp (pemp2 [257.2] from digested plasmids [PCR-product])<br>256. pSBLb4C15-pem (pem1 [258.1] from digested plasmids [PCR-product])<br>256. pSBLb4C15-pem (pem2 [258.2] from digested plasmids [PCR-product])<br>256. pSBLb4C15-pem (pem3 [258.3] from digested plasmids [PCR-product])<br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br><b>Re streak 14/8 or 15/8</b><br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br><br><b>Transformation & plating</b><br>134.1 pSB1C3-CBP (134.1 from digested plasmids [PCR product])<br>134.2 pSB1C3-CBP (134.2 from digested plasmids [PCR product])<br>254.1 pSB1C3-gus (254.1 from digested plasmids [PCR product])<br>254.1 pSB1C3-gus (254.2 from digested plasmids [PCR product])<br>255.2 pSBLb4C15-pemp (257.2 from digest plasmids [PCR product])<br>255.1 pSBLb4C15-pemp (257.1 from digest plasmids [PCR product])256.1 pSBLb4C15-pem (258.1 from digested plasmids [PCR product])<br>256.2 pSBLb4C15-pem (258.2 from digested plasmids [PCR product])<br>225. pEL3S17-J23101-B0034-TAL-B0034-4CL-B0034-STS<br>261. pSB3K3-J23110-B0034-HIS-4CL<br>120. pSB3K3-B0034-HIS-4CL<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>177pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>184.pSB1C3-B0034-CrtZ<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br><br><b>Re-streaking</b><br>254. pSB1C3-gus (gus3 [254.3] from digested plasmids [PCR-product]): 1 clone<br>255. pSBLb4C15-pemp (pemp3 [257.3] from digested plasmids [PCR-product]): 4 clones<br><br><b>O/N:</b><br>275. pSBLb4C15-red (dam-)<br>276. pSBLb4C15-CP1-Exp-B0034-Linker-Miraculin-His (dam)<br>277. pSBLb4C15-CP1-Exp-B0034-Linker-Miraculin-His (dam)<br><br><b>Gradient PCR:</b> <br>CBP (colony PCR from Lb.Reuteri)<br>CSP (colony PCR from Lb.Reuteri)<br>Mcherry (from plasmid preparation)<br>256. pSBLb4C15-pem<br><br><h2>Results</h2><br>One of the two gels from SDS page was succesfull, still have problems with the buffers<br><br><h2>Characterization</h2><br><b>Characterization of Resveratrol production through spectroscopy and HPLC</b><br><br><br><b>Western blot</b><br><b>SDS-page on constructs:</b><br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>115. pSB3K3-J23101-B0034-His-TAL<br><br><br><b>Electro transfer over night</b><br><br><h2>Other experiments</h2><br><b>Casting LB-agar plates:</b><br>Chloramphenicol: 85<br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013814') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-08-14</h1><br><b>Name of participants: </b>Christoffer A, Stephanie H, Alona N, Mikael S, Viktor B, Viktor T, Alexander, Malin, Pontus, Niclas, Christoffer, Nils, Sabri, Hampus.E, Karl H, Ken B, Kristoffer L, Victor S, Marcus H, Theodor L., Peter C, Lovisa P<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>115. pSB3K3-J23101-B0034-His-TAL<br><br><b>E. coli PK401 dam-:</b><br>111.7 pSBLb4C15-red<br>181. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br><br><h2>Todays work:</h2><br><b>Re-streak:</b><br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>110.pEL3S17-K206001-B0034-IspA<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>pSB1K3-J23110-B0034-aeBlue (should be subcloned to pSB1C3 after)<br>pSB3K3-J23110-B0034-AmilCP (should be subcloned to pSB1C3 after)<br>pSB3K3-J23110-B0034-asCP (should be subcloned to pSB1C3 after)<br>Re streak 15/8 or 16/8<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br><br><b>O/N:</b><br>285. pSB1C3-J23110-B0034-aeBlue<br>286. pSB1C3-J23110-B0034-amilCP<br>287. pSB1C3-J23110-B0034-asCP<br><br><b>Streak from frozen stock:</b><br>111. pSBLb4C15-red<br><br><b>Gradient touchdown colony PCR:</b><br>CSP <br><br><b>Gel Electrophoresis:</b><br>CBP (colony PCR from Lb.Reuteri)<br>CSP (colony PCR from Lb.Reuteri)<br>Mcherry (from plasmid preparation)<br>256. pSBLb4C15-pem<br><br><b>PCR purification: </b><br>M1 <br>M2<br>M3<br>M4<br>CB1<br>CB2<br>256. pSBLb4C15-pem (P1)<br><br><b>Digestion:</b><br>M3 (x+p)<br>CB1 (x+p)<br>P1 (x+p)<br>111.7 pSB4C15-red (e+p)<br>Bleomycin resistance cassette (SacI)<br>pEL PCR product with e. coli ori (ClaI)<br>Lb3 reuteri DSM 20016 pVS2 (ClaI)<br>CP promoters with amilGFP, aeBlue,tsPurple<br><br><b>Ligation:</b><br>M3 with pSB1C3, pSBLB4C15 + CP8, CP1<br>CB1 with pSB1C3<br>P1 with pSB1C3<br>pVS2 origin of replication with pEL PCR-product, cut by ClaI <br>CP promoters with amilGFP, aeBlue,tsPurple<br><br><b>Frozen stock and plasmid preparation:</b><br>275.1 pSBLb4C15-red (dam-)*<br>275.2 pSBLb4C15-red (dam-)<br>276. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His (dam-)<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His (dam-)<br><i>*O/N of clone 1 was made from a very small colony, its colour wasn’t as strong as clone 2’s after growing overnight.</i><br><br><b>Transformation:</b><br>109.pEL3S17-K206000-B0034-IspA x<br>110.pEL3S17-K206001-B0034-IspA x<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His x<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His x<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His x<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His x<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His x<br>184.pSB1C3-B0034-CrtZ x<br>208.pSB1A3-B0034-Thioredoxin-ZCD-His x<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His x<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His x<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His x<br>267.psB3K3-K206000-B0034-IspA x<br>268.psB3K3-K206001-B0034-IspA x<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His x<br>272 pSB1K3-B0034-Thioredoxin-ZCD-His x<br>273 pSB3T5-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His x<br>274 pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His x<br>179 pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His x<br><br><b>Plasmid prepp:</b><br>280. pSB1A3-B0034-aeBlue (should be subcloned to pSB1C3 after, give Erik the subclones version)<br><br><h2>Results:</h2><br><b>Gel Electrophoresis:</b><br>CBP (colony PCR from Lb.Reuteri)<br>CB1: 54.3o C (NC ok!), CB2: 63.3o C (NC ok!) → Correct bands!!<br>CSP (colony PCR from Lb.Reuteri) <br>CS1: 55.7o C (NC ok!), CS2: 65.6o C (NC ok!) → No bands!! (not even primer dimer) <br>Mcherry (from plasmid preparation)<br>M1: 45.0o C (NC ok!), M2: 47.9o C (NC ok!), M3: 50.5o C (NC ok!), M4: 54.3o C (NC ok!) → Correct bands!!<br>256. pSBLb4C15-pem: <br>P1: 47.9o C (NC ok!) → Correct bands!!<br><br><b>PCR purification: </b><br>M1: 65.9 ng/µl → Store in freezer!<br>M2: 200.6 ng/µl → Store in freezer!<br>M3: 136.8 ng/µl → For digest! → Store in freezer!<br>M4: 49.5 ng/µl → Store in freezer!<br>CB1: 90.8 ng/µl → For digest! → Store in freezer!<br>CB2: 56.7 ng/µl → Store in freezer!<br>P1: 56.2 ng/µl → For digest! → Store in freezer!<br><br><b>Plasmid preparation:</b><br>275.1 PK401 dam- pSBLb4C15-red (dam-): 13.5 ng/µl<br>275.2 PK401 dam- pSBLb4C15-red (dam-): 27.9 ng/µl<br>276. PK401 dam- pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His (dam-): 18.3 ng/µl<br>277. PK401 dam- pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His (dam-): 26.6 ng/µl<br><br><b>Western blot:</b><br>Transfer overnight not complete but continued with anyway<br>Very promising results, bands can be seen around 50 kDa but the ladder is pretty weak so we can’t be absolutely sure. Looks like the J23101 promotor does’nt work.<br><br><h2>Characterization experiments</h2><br><b>Characterization of Resveratrol production through spectroscopy and HPLC</b><br><b>Blotting and development of constructs:</b><br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>115. pSB3K3-J23101-B0034-His-TAL<br><br><h2>Other experiments:</h2><br><b>Making MRS-broth (2x600 ml)</b><br><br><b>New buffers for western blot made </b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013815') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-08-15</h1><br><b>Name of participants: </b> Christoffer Ahlström, Stephanie Herman, Alona Nyberg, Mikael Strandgren, Alexander, Malin, Pontus, Niclas, Christoffer, Sabri, Hampus.E, Karl H, Ken B, Kristoffer L, Victor S, Marcus H., Peter C, Lovisa P, Theodor L<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>282.pSB3K3-J23110-B0034-amilCP<br>283.pSB3K3-J23110-B0034-asCCP<br><br><b>E. coli PK401 dam-:</b><br>275.2 pSBLb4C15-red<br>276 pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>292. pSBLb4C15-CP1-mCherry<br>294. pSBLb4C15-CP8-mCherry<br><br><h2>Todays work:</h2><br><b>Digestions:</b><br>Erythromycin resistance cassette from pLUL631 (SacI, Tango buffer)<br>Bleomycin resistance cassette (SalI, FD buffer)<br><br><b>O/N Screening glycerolstock:</b><br>110.pEL3S17-K206001-B0034-IspA<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br><br><b>Re-streak:</b><br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>184.pSB1C3-B0034-CrtZ<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>110.pEL3S17-K206001-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br><br><b>Plasmid prep:</b><br>282.pSB3K3-J23110-B0034-amilCP<br>283.pSB3K3-J23110-B0034-asCCP<br><br><b>Transformation:</b><br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br>292. PK401 dam- pSBLb4C15-CP1-mCherry (dam-)<br>294. PK401 dam- pSBLb4C15-CP8-mCherry (dam-)<br><br><b>Ligations:</b><br>pSBLB4C15 and Bleomycin resistance cassette cut with SacI and SalI<br><br><b>Spreading of strains:</b><br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br><i>E. coli PK401 dam-:</i><br>292. pSBLb4C15-CP1-mCherry<br>294. pSBLb4C15-CP8-mCherry<br><br><b>Screening PCR and gel electrophoresis</b><br><br><h2Results:</h2><br><b>Yesterdays buffers for western blot resulted in smear. </b><br><br><h2>Characterization experiments</h2><br><b>Characterization of Resveratrol production through spectroscopy and HPLC</b><br><br><h2>Other experiments</h2><br><b>Making MRS-agar plates:</b><br>47 Chloramphenicol<br>47 Without antibiotics<br><br><b>Testing yesterdays buffers for western blot.</b><br> <br><b>New separation- and stacking buffers for western blot with a factor 1000 concentration of Tris.</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013816') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-08-16</h1><br><b>Name of participants: </b>Anton Berglund, Stephanie Herman, Mikael Strandgren, Alexander, Pontus, Nils, Christoffer, Sabri, Hampus.E, Kristoffer L, Victor S, Marcus H, Karl H, Theodor L, Peter C<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>267 pSB1C3-pemp<br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>282.pSB3K3-J23110-B0034-amilCP<br>283.pSB3K3-J23110-B0034-asCCP<br>225: pEL3S17-J23101-B0034-TAL-B0034-4CL-B0034-STS<br>295: pSB1C3-J23110-TAL-B0034-4Cl-B0034-STS<br><br><b>E. coli PK401 dam-:</b><br>275.2 pSBLb4C15-red<br>276 pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>292. pSBLb4C15-CP1-mCherry<br>294. pSBLb4C15-CP8-mCherry<br><br><h2>Todays work:</h2><br><b>O/N Screening & glycerolstock:</b><br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>184.pSB1C3-B0034-CrtZ<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>2 74.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br><br><b>Streak from frozen stock:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br><br><b>Digestion</b><br>Erythromycin cassette amplicon from pLUL631 (SalI)<br><br><b>Ligation</b><br>pSB1C3-pemp<br>298. pSBLb4E15-red (E-cassette amplicon from pLUL631 with pSBLb4C15 (SacI + SalI))<br>297. pSBLb4B15-red (Bleomycin cassette with pSBLb4C15 (SacI + SalI))<br><br><b>Plasmid preparation:</b><br>204.1 pEL3S17-J23110-B0034-HIS-STS<br>204.3 pEL3S17-J23110-B0034-HIS-STS<br>204.4 pEL3S17-J23110-B0034-HIS-STS<br>254.1 pSB1C3-gus<br>254.2 pSB1C3-gus<br>255.1 pSBLB4C15-pemp<br>255.2 pSBLB4C15-pemp<br>255.3 pSBLB4C15-pemp<br>255.4 pSBLB4C15-pemp<br><br><b>O/N:</b><br>204.1 pEL3S17-J23110-B0034-HIS-STS<br>204.3 pEL3S17-J23110-B0034-HIS-STS<br>204.4 pEL3S17-J23110-B0034-HIS-STS<br>254.1 pSB1C3-gus<br>254.2 pSB1C3-gus<br>255.1 pSBLB4C15-pemp<br>255.2 pSBLB4C15-pemp<br>255.3 pSBLB4C15-pemp<br>255.4 pSBLB4C15-pemp<br>257.1 pSB1C3-pemp<br>257.2 pSB1C3-pemp<br>257.3 pSB1C3-pemp<br>280. pSB1K3-J23110-B0034-aeBlue<br>281. pSB3K3-J23110-B0034-amilCP<br>282. pSB3K3-J23110-B0034-asCP<br><br><b>Transformation:</b><br>297. PK401 dam- pSBLb4B15-red <br>109.pEL3S17-K206000-B0034-IspA <br>110.pEL3S17-K206001-B0034-IspA <br>155pSB1C3-j23110-RBS-meffRed<br>156pSB1C3-j23106-RBS-meffRed<br>157pSB1C3-j23110-RBS-spisPink<br>158pSB1C3-j23106-RBS-spisPink<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His <br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His <br>179 pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His <br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His <br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His <br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His <br>184.pSB1C3-B0034-CrtZ<br>202pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208.pSB1A3-B0034-Thioredoxin-ZCD-His <br>247pSB1C3-J23110-RBS-tsPurple<br>248pSB1C3-J23106-RBS-tsPurple<br>249pSB1C3-J23110-RBS-scOrange<br>250pSB1C3-J23106-RBS-scOrange<br>251pSB1C3-J23110-RBS-fwYellow<br>252pSB1C3-J23106-RBS-fwYellow<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His <br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His <br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His <br>267.psB3K3-K206000-B0034-IspA <br>268.psB3K3-K206001-B0034-IspA <br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His <br>272 pSB1K3-B0034-Thioredoxin-ZCD-His <br>273 pSB3T5-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His <br>274 pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His <br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br>225: pEL3S17-J23101-B0034-TAL-B0034-4CL-B0034-STS<br>295: pSB1C3-J23110-TAL-B0034-4Cl-B0034-STS<br><br><h2>Results:</h2><br><b>Plasmid prep:</b><br>204.1 pEL3S17-J23110-B0034-HIS-STS: 9.7 ng/µl<br>204.3 pEL3S17-J23110-B0034-HIS-STS: 11.9 ng/µl<br>204.4 pEL3S17-J23110-B0034-HIS-STS: 7.6 ng/µl<br>254.1 pSB1C3-gus: 6.7 ng/µl<br>254.2 pSB1C3-gus: 11.5 ng/µl<br>255.1 pSBLB4C15-pemp: 9.9 ng/µl<br>255.2 pSBLB4C15-pemp: 6.3 ng/µl<br>255.3 pSBLB4C15-pemp: 5.0 ng/µl<br>255.4 pSBLB4C15-pemp: 4.6 ng/µl<br><i>Since the concentrations were so low a new set of overnight cultures were prepared.</i><br><br><b>Spectophotometry:</b><br>Inconclusive results.<br><br><h2>Characterization experiments</h2><br><b>Characterization of Resveratrol production through spectroscopy and HPLC</b><br><br><b>Testing of new buffers for western blot.</b><br><br><h2>Other experiments</h2><br><b>Making MRS-broth (2x600 ml)</b><br><br><b>Making LB-agar plates:</b><br>173 Chloramphenicol <i>(chloramphenicol was forgotten in 26 of the plates, but they had already been mixed with 25 others, so also these had to be discarded. 5 plates of doubtful antibiotic content had already been used by the carotenoid group, they were probably ones with ab. E. colis containing pSB3K3 were spread on some of the mixed plates.)</i><br>28 Kanamycin</div>' | ||
| + | } | ||
| + | else if(id == 'd2013817') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-08-17</h1><br><b>Name of participants: </b>Anders Edlund, Alexander, Pontus, Sabri, Hampus.E, Kristoffer L, Viktor @, Marcus H. Ken B-A, Karl H, Theodor L<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>257. pSB1C3-pemp<br>267 pSB1C3-pemp<br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry 58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>282.pSB3K3-J23110-B0034-amilCP<br>283.pSB3K3-J23110-B0034-asCCP<br>299.pEL3S17-J23101-B0034-Thioredoxin<br>300.pEL3S17-J23106-B0034-Thioredoxin<br>301.pEL3S17-J23110-B0034-Thioredoxin<br>225: pEL3S17-J23101-B0034-TAL-B0034-4CL-B0034-STS<br>295: pSB1C3-J23110-TAL-B0034-4Cl-B0034-STS<br><br><b>E. coli PK401 dam-:</b><br>275.2 pSBLb4C15-red<br>276 pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>292. pSBLb4C15-CP1-mCherry<br>294. pSBLb4C15-CP8-mCherry<br><br><h2>Todays work:</h2><br><b>Restreak</b><br>254.3 pSB1C3-Gus<br>288.1 pSB1C3-pem<br>288.2 pSB1C3-pem<br>288.3 pSB1C3-pem<br>288.4 pSB1C3-pem<br>289.1 pSB1C3-mCherry<br>289.2 pSB1C3-mCherry<br>289.3 pSB1C3-mCherry<br>289.4 pSB1C3-mCherry<br>297. pSBLb4B15-red <br>202pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>207pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1 <br>225: pEL3S17-J23101-B0034-TAL-B0034-4CL-B0034-STS<br>295: pSB1C3-J23110-TAL-B0034-4Cl-B0034-STS<br><br><b>Plasmid preparation:</b><br>254.1 pSB1C3-Gus<br>254.2 pSB1C3-Gus<br>255.1 pSBLB4C15-pemp<br>255.2 pSBLB4C15-pemp<br>257.1 pSB1C3-pemp<br>257.2 pSB1C3-pemp<br>184.pSB1C3-B0034-CrtZ<br>280. pSB1K3-J23110-B0034-aeBlue<br>281. pSB3K3-J23110-B0034-amilCP<br>282. pSB3K3-J23110-B0034-asCP<br><br><b>Digest:</b><br>147.pSB1C3-B0034-Thioredoxin (X,P)<br><br><b>Ligation:</b><br>147.pSB1C3-B0034-Thioredoxin, 91.BBa_J61002 (amp)-J23106, 36.pEL3S17-red<br>---> 300.pEL3S17-J23106-B0034-Thioredoxin<br>147.pSB1C3-B0034-Thioredoxin, 92.BBa_J61002 (amp)-J23110, 36.pEL3S17- red<br>---> 301.pEL3S17-J23110-B0034-Thioredoxin<br>147.pSB1C3-B0034-Thioredoxin, 103.pSB1K3-J23101, 36.pEL3S17-red<br>---> 299.pEL3S17-J23101-B0034-Thioredoxin<br><br><b>Transformation:</b><br>203pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>299.pEL3S17-J23101-B0034-Thioredoxin<br>300.pEL3S17-J23106-B0034-Thioredoxin<br>301.pEL3S17-J23110-B0034-Thioredoxin<br><br><h2>Results:</h2><br><b>Plasmid preparation:</b><br>254.1 pSB1C3-Gus 110 ng/uL<br>254.2 pSB1C3-Gus 132 ng/uL<br>255.1 pSBLB4C15-pemp156 ng/uL<br>255.2 pSBLB4C15-pemp 158 ng/uL<br>257.1 pSB1C3-pemp 146 ng/uL<br>257.2 pSB1C3-pemp 140 ng/uL</div>' | ||
| + | } | ||
| + | else if(id == 'd2013818') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Sunday 2013-08-18</h1><br><b>Name of participants: </b>Anders Edlund, Alexander, Pontus, Kristoffer L, Marcus H.<br><br><h2>Ongoing constructs:</h2><br><h2>E. coli:</h2><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>267 pSB1C3-pemp<br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>282.pSB3K3-J23110-B0034-amilCP<br>283.pSB3K3-J23110-B0034-asCCP<br>299.pEL3S17-J23101-B0034-Thioredoxin<br>300.pEL3S17-J23106-B0034-Thioredoxin<br>301.pEL3S17-J23110-B0034-Thioredoxin<br><br><b>E. coli PK401 dam-:</b><br>275.2 pSBLb4C15-red<br>276 pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>292. pSBLb4C15-CP1-mCherry<br>294. pSBLb4C15-CP8-mCherry<br><br><h2>Todays work: </h2><br><b>ON:</b><br>134. pSB1C3-B0034-CBP<br>254. pSB1C3-gus<br>288.1 pSB1C3-pem<br>288.2 pSB1C3-pem<br>288.3 pSB1C3-pem<br>288.4 pSB1C3-pem<br>289.1 pSB1C3-mCherry<br>289.2 pSB1C3-mCherry<br>289.3 pSB1C3-mCherry<br>289.4 pSB1C3-mCherry<br>pSBLB4C15-red<br>pSBLB4C15-red dam-<br><br><b>Restreak:</b><br>68.1 pSB3K3-B0034-BFP<br>68.2 pSB3K3-B0034-BFP<br>68.4 pSB3K3-B0034-BFP<br>109.pEL3S17-K206000-B0034-IspA <br>110.pEL3S17-K206001-B0034-IspA <br>147.pSB1C3-B0034-Thioredoxin<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His <br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His <br>179 pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His <br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His <br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His <br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His <br>265.pSB3K3-J23110-B0034-Exp-Linker-Miraculin-His <br>266.pSB3K3-CP1-B0034-Exp-Linker-Miraculin-His <br>267.pSB3K3-K206000-B0034-IspA <br>268.pSB3K3-K206001-B0034-IspA <br>269.pSB3K3-CP8-B0034-Exp-Linker-Miraculin-His <br>300.pEL3S17-J23106-B0034-Thioredoxin<br><br><b>PCR:</b> <br>134. pSB1C3-B0034-CBP<br>254. pSB1C3-gus<br>288.1 pSB1C3-pem<br>288.2 pSB1C3-pem<br>288.3 pSB1C3-pem<br>288.4 pSB1C3-pem<br>289.1 pSB1C3-mCherry<br>289.2 pSB1C3-mCherry<br>289.3 pSB1C3-mCherry<br>289.4 pSB1C3-mCherry<br><br><b>Digest:</b><br>125. pSB1C3-ZCD-His<br>127. pSB1A3-B0034-UGTCs2-His<br>129.pSB1C3-Idi(mut) <br>184.pSB1C3-B0034-CrtZ<br><br><b>Ligation:</b><br>4.pSB1A3-red<br>7.pSB3K3-red<br>36.pEL3S17-red<br>91.j23106<br>92.j23110<br>103.pSB1K3-j23101<br>125. pSB1C3-ZCD-His<br>127. pSB1A3-B0034-UGTCs2-His<br>129.pSB1C3-Idi(mut) <br>184.pSB1C3-B0034-CrtZ<br><br><b>Plasmidprep:</b><br>184.pSB1C3-B0034-CrtZ<br><br><b>Transformation:</b><br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>203pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>302. idi (mut)-B0034-Ispa<br>303.pSB1A3-j23101-B0034-CrtZ<br>304.pSB1A3-j23106-B0034-CrtZ<br>305.pSB1A3-j23110-B0034-CrtZ<br><br><b>O/N screening:</b><br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br><br><h2>Results:</h2><br><b>Failed:</b><br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His <br>184.pSB1C3-B0034-CrtZ<br>202pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208.pSB1A3-B0034-Thioredoxin-ZCD-His <br>272 pSB1K3-B0034-Thioredoxin-ZCD-His <br>273 pSB3T5-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His <br>274 pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His</div>' | ||
| + | } | ||
| + | else if(id == 'd2013819') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-08-19</h1><br><b>Name of participants: </b>Anton Berglund, Viktor Blomkvist, Mikael Strandgren, Stephanie Herman, Kristoffer L, Marcus H, Karl H, Theodor L, Emil M, Victor S, Alexander, Pontus, Sabri<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>257. pSB1C3-pemp<br>267 pSB1C3-pemp<br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>282.pSB3K3-J23110-B0034-amilCP<br>283.pSB3K3-J23110-B0034-asCCP<br>299.pEL3S17-J23101-B0034-Thioredoxin<br>300.pEL3S17-J23106-B0034-Thioredoxin<br>301.pEL3S17-J23110-B0034-Thioredoxin<br><br><b>E. coli PK401 dam-:</b><br>275.2 pSBLb4C15-red<br>276 pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>292. pSBLb4C15-CP1-mCherry<br>294. pSBLb4C15-CP8-mCherry<br><br><h2>Todays work: </h2><br><b>Screening</b><br>134.1 pSB1C3-CBP<br>254 pSB1C3-gus<br>288.1 pSB1C3-pem<br>288.2 pSB1C3-pem<br>288.3 pSB1C3-pem<br>288.4 pSB1C3-pem<br>289.1 pSB1C3-mCherry<br>289.2 pSB1C3-mCherry<br>289.3 pSB1C3-mCherry<br>289.4 pSB1C3-mCherry<br><br><b>Screening O/N:</b><br>109.pEL3S17-K206000-B0034-IspA <br>110.pEL3S17-K206001-B0034-IspA <br>147.B0034-Thioredoxin<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His <br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His <br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His <br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His <br>267.psB3K3-K206000-B0034-IspA <br>268.psB3K3-K206001-B0034-IspA <br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His <br>300.J23106-B0034-Thioredoxin<br>202.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br><br><b>Digestion & screening:</b><br>254.1 pSB1C3-gus: XbaI & PstI<br>254.2 pSB1C3-gus: XbaI & PstI<br>255.1 pSBLB4C15-pemp: EcoRI & PstI<br>255.2 pSBLB4C15-pemp: EcoRI & PstI<br>257.1 pSB1C3-pemp: EcoRI & PstI<br>257.2 pSB1C3-pemp: EcoRI & PstI<br><br><b>Frozen stock:</b><br>134.1 pSB1C3-CBP<br>275. pSBLB4C15-red (dam-)<br>288.1 pSB1C3-pem<br>288.2 pSB1C3-pem<br>288.4 pSB1C3-pem<br>289.1 pSB1C3-mCherry<br>289.3 pSB1C3-mCherry<br>289.4 pSB1C3-mCherry<br><br><b>1 ml overnight culture:</b><br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>139. pSB3K3-J23101-B0034-BFP<br><br><b>Plasmid preparation</b><br>111.7(1) pSBLB4C15-red<br>111.7(2) pSBLB4C15-red<br>134.1 pSB1C3-CBP<br>275 pSBLB4C15-red (dam-)<br>288.1 pSB1C3-pem<br>288.2 pSB1C3-pem<br>288.4 pSB1C3-pem<br>289.1 pSB1C3-mCherry<br>289.3 pSB1C3-mCherry<br>289.4 pSB1C3-mCherry<br><br><b>Overnight culture:</b><br>Lb11.2 plantarum 36E-no plasmid<br><br><b>Phusion PCR:</b><br>usp45 (synthesized signal peptide from Lactococcus lactis)<br><br><b>Restreak</b><br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br><br><b>digest</b><br>36. pEL3S17-red<br><br><b>Ligation</b><br>108.pEL3S17-J23106-B0034-IspA<br>140.pEL3S17-J23110-B0034-UGTCs2-His<br>141.pEL3S17-J23101-B0034-UGTCs2-His<br>142.pEL3S17-CP1-B0034-UGTCs2-His<br>143.pEL3S17-CP8-B0034-UGTCs2-His<br><br><b>Subclone:</b><br>280. pSB1A3-B0034-aeBlue à 284. pSB1C3-B0034-aeBlue<br>281. pSB1K3-J23110-B0034-aeBlue à 285. pSB1C3-J23110-B0034-aeBlue<br>282. pSB3K3-J23110-B0034-amilCP à 286. pSB1C3-J23110-B0034-amilCP<br>283. pSB3K3-J23110-B0034-asCP à 287. pSB1C3-J23110-B0034-asCP<br><br><b>Two step Assembly:</b><br>92. BBa_J61002-J23110, 149. pSB1C3-RBS-meffRed à 155. pSB1C3-J23110-RBS-meffRed<br>91. BBa_J61002-J23106, 149. pSB1C3-RBS-meffRed à 156. pSB1C3-J23106-RBS-meffRed<br>92. BBa_J61002-J23110, 150. pSB1C3-RBS-spisPink à 157. pSB1C3-J23110-RBS-spisPink<br>91. BBa_J61002-J23106, 150. pSB1C3-RBS-spisPink à 158.pSB1C3-J23106-RBS-spisPink<br>92. BBa_J61002-J23110, 148. pSB1C3-RBS-gfasPurple à 159.pSB1C3-J23110-RBS-gfasPurple<br>91. BBa_J61002-J23106, 148 pSB1C3-RBS-gfasPurple à 160.pSB1C3-J23106-RBS-gfasPurple<br><br><h2>Results:</h2><br><b>Screening</b><br>134.1 pSB1C3-CBP: showed the correct band!<br>254 pSB1C3-gus: showed a very weak band, not sure if correct!<br>288.1 pSB1C3-pem: showed the correct band!<br>288.2 pSB1C3-pem: showed the correct band!<br>288.3 pSB1C3-pem: did not show the correct band!<br>288.4 pSB1C3-pem: showed the correct band!<br>289.1 pSB1C3-mCherry: showed the correct band!<br>289.2 pSB1C3-mCherry: did not show the correct band!<br>289.3 pSB1C3-mCherry: showed the correct band!<br>289.4 pSB1C3-mCherry: showed the correct band.<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His: fail<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His: fail<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His: fail<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His: fail<br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268: fail<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1: fail<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1: fail<br><br><b>Digestion & screening:</b><br>254.1 pSB1C3-gus: XbaI & PstI, did not show the correct bands!<br>254.2 pSB1C3-gus: XbaI & PstI, did not show the correct bands!<br>255.1 pSBLB4C15-pemp: EcoRI & PstI, showed the correct bands!<br>255.2 pSBLB4C15-pemp: EcoRI & PstI, showed the correct bands!<br>257.1 pSB1C3-pemp: EcoRI & PstI, showed the correct bands!<br>257.2 pSB1C3-pemp: EcoRI & PstI, showed the correct bands!<br><br><b>Plasmid preparation</b><br>111.7(1) pSBLB4C15-red: 81.7 ng/µl<br>111.7(2) pSBLB4C15-red: 93.9 ng/µl<br>134.1 pSB1C3-CBP: 163.8 ng/µl<br>275 pSBLB4C15-red (dam-): 92.4 ng/µl<br>288.1 pSB1C3-pem: 211.2 ng/µl<br>288.2 pSB1C3-pem: 193.2 ng/µl<br>288.4 pSB1C3-pem: 193.9 ng/µl<br>289.1 pSB1C3-mCherry: 230.4 ng/µl<br>289.3 pSB1C3-mCherry: 229.7 ng/µl<br>289.4 pSB1C3-mCherry: 239.7 ng/µl</div>' | ||
| + | } | ||
| + | else if(id == 'd2013820') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-08-20</h1><br><b>Name of participants: </b>Anton Berglund, Stephanie Herman, Alona Nyberg, Alexander, Pontus, Sabri, Malin, Magnus, Marcus H, Karl H, Theodor L, Emil M, Victor S<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>257. pSB1C3-pemp<br>267 pSB1C3-pemp<br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>299.pEL3S17-J23101-B0034-Thioredoxin<br>300.pEL3S17-J23106-B0034-Thioredoxin<br>301.pEL3S17-J23110-B0034-Thioredoxin<br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>(201. E.Coli. Nissle - CrtY-CrtI)<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br>147.pSB1C3-B0034-Thioredoxin<br>184.pSB1C3-B0034-CrtZ<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br><br><b>E. coli PK401 dam-:</b><br>275.2 pSBLb4C15-red<br>276 pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>292. pSBLb4C15-CP1-mCherry<br>294. pSBLb4C15-CP8-mCherry<br><br><h2>Todays work: </h2><br><b>Characterisation of the CP promoters with facs:</b><br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br><br><b>Digestion:</b><br>USP. with E, P<br>191. pSB1A2-fusionGFP with X, P<br>280. pSB1A3-B0034-aeBlue with X, P <br>147.B0034-Thioredoxin (E, S)<br>147.B0034-Thioredoxin (X,P) <br>27.pEL3K16-red (E,P)<br>125.pSB1C3-ZCD-His (X,P)<br><br><b>Ligation:</b><br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>299.pEL3S17-J23101-B0034-Thioredoxin<br>300.pEL3S17-J23106-B0034-Thioredoxin<br>301.pEL3S17-J23110-B0034-Thioredoxin<br><br><b>Transformation:</b><br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br><br><b>Screen/ ON:</b><br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>303.pSB1A3-j23101-B0034-CrtZ<br>304.pSB1A3-j23106-B0034-CrtZ<br><br><b>Plasmid prep. / Seq. prep.</b><br>147.pSB1C3-B0034-Thioredoxin<br>184.pSB1C3-B0034-CrtZ<br><br><b>Gel:</b><br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>207.CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br><br><b>Phusion PCR:</b><br>->219.pSB1C3-Miraculin<br><br><b>Making competent Lactobacillus reuteri</b><br>Lb12.1 reuteri DSM 20016 plasmid free: eight<br>Lb12.2 reuteri DSM 20016 plasmid free: eight<br><br><b>Electroporation</b><br>Lb3.2 reuteri DSM 20016 pVS2: two<br>111.7(1) pSBLB4C15-red: one<br>111.7(2) pSBLB4C15-red: two<br>255.1 pSBLB4C15-pemp: one<br>255.2 pSBLB4C15-pemp: one<br>275 pSBLB4C15-red (dam-): three<br><br><h2>Results:</h2><br><b>Electroporation</b><br>Lb3.2 reuteri DSM 20016 pVS2: one of the two survived<br>111.7(1) pSBLB4C15-red: survived<br>111.7(2) pSBLB4C15-red: one of the two survived<br>255.1 pSBLB4C15-pemp: survived<br>255.2 pSBLB4C15-pemp: survived <br>275 pSBLB4C15-red (dam-): two of the three survived</div>' | ||
| + | } | ||
| + | else if(id == 'd2013821') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-08-21</h1><br><b>Name of participants: </b>Anton Berglund, Mikael Strandgren, Viktor Blomkvist, Alexander, Pontus, Sabri, Malin, Magnus, christoffer, Marcus H, Karl H, Theodor L, Emil M, Victor S, Hampus.E, Lovisa P<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>257. pSB1C3-pemp<br>267 pSB1C3-pemp<br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>(201. E.Coli. Nissle - CrtY-CrtI)<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br>204(1-4). pEL3S17-J23110-B0034-His-STS<br><br><b>E. coli PK401 dam-:</b><br>275.2 pSBLb4C15-red<br>276 pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>292. pSBLb4C15-CP1-mCherry<br>294. pSBLb4C15-CP8-mCherry<br><br><h2>Todays work: </h2><br><b>Digestion:</b><br>usp45: XbaI & AgeI<br>191.1 pSB1A2-fusionGFP: XbaI, PstI<br>191.1 pSB1A2-fusionGFP: NgoMIV & PstI<br>275. pSBLB4C15-red (dam-): EcoRI & PstI<br>289.1 pSB1C3-mCherry: NgoMIV & PstI<br>289.1 pSB1C3-mCherry: XbaI, PstI<br>297.1 pSBLB4B15-red (dam-): EcoRI & PstI<br><br><b>Ligation:</b><br>pSBLB4B15-CP11-fusionGFP (297.1+63.2+191.1)<br>pSBLB4B15-CP30-fusionGFP (297.1+65.1+191.1) <br>pSBLB4B15-CP11-usp45-fusionGFP (297.1+63.2+usp45+191.1) <br>pSBLB4B15-CP30-usp45-fusionGFP (297.1+65.2+usp45+191.1) <br>pSBLB4B15-CP11-usp45-mCherry (297.1+63.2+usp45+289.1) <br>pSBLB4B15-CP30-usp45-mCherry (297.1+65.2+usp45+289.1) <br>pSB1C3-usp45 (1+usp45)<br>pSB1C3-usp45 (6+usp45)<br>pSBLb4C15-CP1-B0034-BFP<br>pSBLb4C15-CP8-B0034-BFP<br>pSBLb4C15-CP11-B0034-BFP<br>pSBLb4C15-CP29-B0034-BFP<br>pSBLb4C15-CP30-B0034-BFP<br>pSBLb4C15-CP41-B0034-BFP<br>pSBLb4C15-CP44-B0034-BFP<br>pSBLb4C15-CP11-aeblue<br>pSBLb4C15-CP30-aeblue<br>pSBLb4B15-CP11-mcherry<br>pSBLb4B15-CP30-mcherry<br>pSBLb4B15-CP11-tspurple<br>pSBLb4B15-CP30-tspurple<br>pSBLb4B15-CP11-amilCP<br>pSBLb4B15-CP30-amilCP<br>pSBLb4B15-CP11-amilGFP<br>pSBLb4B15-CP30-amilGFP<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>265.pSB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.pSB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.pSB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>299.pEL3S17-J23101-B0034-Thioredoxin<br>300.pEL3S17-J23106-B0034-Thioredoxin<br>301.pEL3S17-J23110-B0034-Thioredoxin<br>303.pSB1A3-j23101-B0034-CrtZ<br>304.pSB1A3-j23106-B0034-CrtZ<br><br><b>Transformation:</b><br>108.pEL3S17-J23106-B0034-IspA<br>141.pRL3S17-J23101-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>208.pSB1A3-B0034-Thioredoxin-ZCD-His<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>272.pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>303.pSB1A3-j23101-B0034-CrtZ<br>304.pSB1A3-j23106-B0034-CrtZ<br>204(1-4)<br> <br><b>Re-streak:</b><br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br><br><b>Screen/ ON:</b><br>219.pSB1C3-Miraculin<br>304.pSB1A3-J23106-B0034-CrtZ<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>280. pSB1A3-B0034-aeBlue<br>281. pSB1K3-J23110-B0034-aeBlue<br>282. pSB3K3-J23110-B0034-amilCP<br>283. pSB3K3-J23110-B0034-asCP <br><br><b>Plasmid prep. </b><br>110.pEL3S17-K206001-B0034-IspA<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br><br><b>Mutagenesis PCR:</b><br>336. pSB1C3-spisPink<br>337. pSB1C3-scOrange<br>338. pSB1C3-meffBlue<br>339. pSB1C3-gfasPurple<br>340. pSB1C3-amajCP(amajLime)<br>341. pSB1C3-tsPurple<br>342. pSB1C3-fwYellow<br>257. pSB1C3-pemp<br>288. pSB1C3-pem<br><br><h2>Other experiments:</h2><br><b>Phosphorylation of primers (20µl of each):</b><br>Pem-mutE_F<br>Pem-mutE_R<br>Pem-mutS_F<br>Pem-mutS_R<br><br><b>Cleaning of electroporation cuvettes</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013822') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-08-22</h1><br><b>Name of participants: </b>Anders Edlund, Anton Berglund, Mikael Strandgren, Viktor Blomkvist, Stephanie Herman, Alona Nyberg, Alexander, Pontus, Sabri, Malin, Magnus<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>257. pSB1C3-pemp<br>267 pSB1C3-pemp<br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>299.pEL3S17-J23101-B0034-Thioredoxin<br>300.pEL3S17-J23106-B0034-Thioredoxin<br>301.pEL3S17-J23110-B0034-Thioredoxin<br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>(201. E.Coli. Nissle - CrtY-CrtI)<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br>147.pSB1C3-B0034-Thioredoxin<br>184.pSB1C3-B0034-CrtZ<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>115. pSB3K5-J23101-B0034-His-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>119. pSB1C3-B0034-4CL (from 15)<br><br><b>E. coli PK401 dam-:</b><br>275.2 pSBLb4C15-red<br>276 pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>292. pSBLb4C15-CP1-mCherry<br>294. pSBLb4C15-CP8-mCherry<br><br><h2>Todays work:</h2><br><b>Competent cell preparation:</b><br>Lb11.1 plantarum 36E no plasmid<br>Lb12.1 reuteri DSM 20016 no plasmid<br>Lb12.2 reuteri DSM 20016 no plasmid<br><br><b>Digest:</b><br>147.B0034-Thioredoxin (E, S)<br>147.B0034-Thioredoxin (X,P) <br>27.pEL3K16-red (E,P)<br>125.pSB1C3-ZCD-His (X,P)<br><br><b>Ligation:</b><br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>299.pEL3S17-J23101-B0034-Thioredoxin<br>300.pEL3S17-J23106-B0034-Thioredoxin<br>301.pEL3S17-J23110-B0034-Thioredoxin<br><br><b>Transformation:</b><br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>Lb31. plantarum 36E pVS2<br>Lb32. plantarum 36E pSBLB4B15-CP30-fusionGFP<br>Lb33. plantarum 36E pSBLB4B15-CP11-usp45-fusionGFP<br>Lb34. plantarum 36E pSBLB4B15-CP11-tsPurple<br>Lb35. reuteri DSM 20016 pVS2<br>Lb36. reuteri DSM 20016 pSBLB4B15-CP30-mCherry<br>Lb37. reuteri DSM 20016 pSBLB4B15-CP30-amilGFP<br>Lb38. reuteri DSM 20016 pSBLB4B15-CP11-amilCP<br>Lb39. reuteri DSM 20016 pSBLB4B15-CP30-amilCP<br>Lb40. reuteri DSM 20016 pSBLB4C15-CP11-aeBlue<br>Lb41. reuteri DSM 20016 pSBLB4C15-CP30-aeBlue<br>Lb42. reuteri DSM 20016 pSBLB4C15-CP1-BFP<br>Lb43. reuteri DSM 20016 pSBLB4C15-CP8-BFP<br>Lb44. reuteri DSM 20016 pSBLB4C15-CP11-BFP<br>Lb45. reuteri DSM 20016 pSBLB4C15-CP29-BFP<br>Lb46. reuteri DSM 20016 pSBLB4C15-CP30-BFP<br><strike>3.2. pVS2 (died in electroporation)<br>310. pSBLB4B15-CP11-fusionGFP (died in electroporation)<br>318. pSBLB4B15-CP11-mCherry (died in electroporation)<br>320. pSBLB4B15-CP11-amilGFP (died in electroporation)<br>313. pSBLB4B15-CP30-usp45-fusionGFP (died in electroporation)<br>317. pSBLB4B15-CP30-tsPurple (died in electroporation)<br>331. pSBLB4C15-CP41-BFP (died in electroporation)<br>332. pSBLB4C15-CP44-BFP (died in electroporation)</strike><br><b>+ other E. coli transformations</b><br><br><b>Mutagenesis PCR:</b><br>257. pSB1C3-pemp (two tubes) with Pem-mutE_F & Pem-mutE_R<br>288. pSB1C3-pem (two tubes) with Pem-mutE_F & Pem-mutE_R<br><br><b>Screen/ ON:</b><br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>303.pSB1A3-j23101-B0034-CrtZ<br>304.pSB1A3-j23106-B0034-CrtZ<br><br><b>Plasmid prep. / Seq. prep.</b><br>147.pSB1C3-B0034-Thioredoxin<br>184.pSB1C3-B0034-CrtZ<br><br><b>Gel:</b><br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>207.CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br><br><b>Phusion PCR:</b><br>->219.pSB1C3-Miraculin<br><br><h2>Results:</h2><br><b>Mutagenesis PCR:</b><br>257. pSB1C3-pemp: Problem with PCR-machine, paused at 98*C for 25 min. → re-do.<br>288. pSB1C3-pem: --”--<br><br><b>Transformation:</b><br>11.1 Lb. plantarum 36E:<br>3.2 pVS2: survived electroporation<br>3.2 pVS2: died during electroporation<br>310. pSBLB4B15-CP11-fusionGFP: died during electroporation<br>311. pSBLB4B15-CP30-fusionGFP: survived electroporation<br>312. pSBLB4B15-CP11-usp45-fusionGFP: survived electroporation<br>313. pSBLB4B15-CP30-usp45-fusionGFP: died during electroporation<br>316. pSBLB4B15-CP11-tsPurple: survived electroporation<br>317. pSBLB4B15-CP30-tsPurple: died during electroporation<br><br><b>12.2 Lb. reuteri DSM 20016:</b><br>3.2 pVS2: survived electroporation<br>318. pSBLB4B15-CP11-mCherry: died during electroporation<br>319. pSBLB4B15-CP30-mCherry: survived electroporation<br>320. pSBLB4B15-CP11-amilGFP: died during electroporation<br>321. pSBLB4B15-CP30-amilGFP: survived electroporation<br>322. pSBLB4B15-CP11-amilCP: survived electroporation<br>323. pSBLB4B15-CP30-amilCP: survived electroporation<br>324. pSBLB4C15-CP11-aeBlue: survived electroporation<br><br><b>12.1 Lb. reuteri DSM 20016:</b><br>325. pSBLB4C15-CP30-aeBlue: survived electroporation<br>326. pSBLB4C15-CP1-BFP: survived electroporation<br>327. pSBLB4C15-CP8-BFP: survived electroporation<br>328. pSBLB4C15-CP11-BFP: survived electroporation<br>329. pSBLB4C15-CP29-BFP: survived electroporation<br>330. pSBLB4C15-CP30-BFP: survived electroporation<br>331. pSBLB4C15-CP41-BFP: died during electroporation<br>332. pSBLB4C15-CP44-BFP: died during electroporation<br><br><h2>Characterization:</h2><br><b>Western blot analysis (SDS-page) on constructs:</b><br>115. pSB3K5-J23101-B0034-His-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>119. pSB1C3-B0034-4CL (from 15) (neg.contr)<br><br><h2>Other experiments:</h2><br><b>Phosphorylation of primers (60 µl of each):</b><br>Pem-mutE_F<br>Pem-mutE_R<br>Pem-mutS_F<br>Pem-mutS_R</div>' | ||
| + | } | ||
| + | else if(id == 'd2013823') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-08-23</h1><br><b>Name of participants: </b>Anton Berglund, Stephanie Herman, Viktor Berglund, Mikael Strandgren, Anders Edlund, Alexander, Pontus, Sabri, Malin, Magnus, Marcus H, Karl H, Hampus.E, Lovisa P, Kristoffer L, Emil M<br><br><h2>Ongoing constructs:</h2><br><b>E. coli (D5α):</b><br>111.7 pSBLb4C15-red<br>257. pSB1C3-pemp (two tubes) with Pem-mutE_F & Pem-mutE_R<br>288. pSB1C3-pem (two tubes) with Pem-mutE_F & Pem-mutE_R<br>102.pSB1C3-CrtE<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>(201. E.Coli. Nissle - CrtY-CrtI)<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br>305.pSB1K3-J23110-CrtZ<br>308.pSB1C3-Miraculin<br>309.pSB1C3-B0034-Miraculin<br>261. pSB3K3-J23110-B0034-His-4CL<br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br><br><h2>Todays work:</h2><br><b>Gel electrophoresis:</b><br>257. pSB1C3-pemp (two tubes) with Pem-mutE_F & Pem-mutE_R<br>288. pSB1C3-pem (two tubes) with Pem-mutE_F & Pem-mutE_R<br>111.7 pSBLb4C15-red (cut with E and P) (to get reference/examine cutsites)<br><br><b>Mutagenesis PCR:</b><br>257. pSB1C3-pemp (two tubes) with Pem-mutE_F & Pem-mutE_R<br>288. pSB1C3-pem (two tubes) with Pem-mutE_F & Pem-mutE_R<br>Touchdown with Tm1+2 and Tm1+4.<br><br><b>Transformation:</b><br>102.pSB1C3-CrtE<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br>305.pSB1K3-J23110-CrtZ<br>308.pSB1C3-Miraculin<br>309.pSB1C3-B0034-Miraculin<br><br><b>Re-streak:</b><br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>339. pSB1C3-gfasPurple<br>340. pSB1C3-amajCP(amajLime)<br>341. pSB1C3-tsPurple<br>342. pSB1C3-fwYellow<br><br><b>Screening O/N:</b><br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>301. pEL3S17-J23110-B0034-Thioredoxin<br><br><b>O/N:</b><br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br><br><b>Subclone:</b><br>280. pSB1A3-B0034-aeBlue à 284. pSB1C3-B0034-aeBlue<br>281. pSB1K3-J23110-B0034-aeBlue à 285. pSB1C3-J23110-B0034-aeBlue<br>282. pSB3K3-J23110-B0034-amilCP à 286. pSB1C3-J23110-B0034-amilCP<br>283. pSB3K3-J23110-B0034-asCP à 287. pSB1C3-J23110-B0034-asCP<br><br><b>Glycerol stock Plasmidprep/sekvensprep:</b><br>108.pEL3S17-J23106-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>184.pSB1C3-B0034-CrtZ<br>299.pEL3S17-J23101-B0034-Thioredoxin<br><br><h2>Results:</h2><br><b>Gel electrophoresis:</b><br>257. pSB1C3-pemp with Pem-mutE_F & Pem-mutE_R: Unspecific binding, nc ok!*<br>288. pSB1C3-pem with Pem-mutE_F & Pem-mutE_R: Unspecific binding, nc ok!*<br>111.7 pSBLb4C15-red (cut with E and P): No band<br><i> *→ redo mutagenesis PCR with higher temperatures</i></div>' | ||
| + | } | ||
| + | else if(id == 'd2013824') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-08-24</h1><br><b>Name of participants: </b>Mikael Strandgren, Anders Berglund, Alexander, Pontus, Sabri, Magnus, Marcus H, Karl H, Hampus.E, Lovisa P, Kristoffer L, Emil M<br><br><h2>Ongoing constructs:</h2><br><b>Escherichia coli</b><br>257. pSB1C3-pemp<br>288. pSB1C3-pem<br>136. pSB1C3-B0034-USP<br>pSB1C3-B0034-usp45<br>102.pSB1C3-CrtE<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>(201. E.Coli. Nissle - CrtY-CrtI)<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br>305.pSB1K3-J23110-CrtZ<br>308.pSB1C3-Miraculin<br>309.pSB1C3-B0034-Miraculin<br>261. pSB3K3-J23110-B0034-His-4CL<br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br><br><b>Lactobacillus reuteri:</b><br>Lb35. reuteri DSM 20016 pVS2<br>Lb36. reuteri DSM 20016 pSBLB4B15-CP30-mCherry<br>Lb37. reuteri DSM 20016 pSBLB4B15-CP30-amilGFP<br>Lb38. reuteri DSM 20016 pSBLB4B15-CP11-amilCP<br>Lb39. reuteri DSM 20016 pSBLB4B15-CP30-amilCP<br>Lb40. reuteri DSM 20016 pSBLB4C15-CP11-aeBlue<br>Lb41. reuteri DSM 20016 pSBLB4C15-CP30-aeBlue<br>Lb42. reuteri DSM 20016 pSBLB4C15-CP1-BFP<br>Lb43. reuteri DSM 20016 pSBLB4C15-CP8-BFP<br>Lb44. reuteri DSM 20016 pSBLB4C15-CP11-BFP<br>Lb45. reuteri DSM 20016 pSBLB4C15-CP29-BFP<br>Lb46. reuteri DSM 20016 pSBLB4C15-CP30-BFP<br><br><b>Lactobacillus plantarum:</b><br>Lb31. plantarum 36E pVS2<br>Lb32. plantarum 36E pSBLB4B15-CP30-fusionGFP<br>Lb33. plantarum 36E pSBLB4B15-CP11-usp45-fusionGFP<br>Lb34. plantarum 36E pSBLB4B15-CP11-tsPurple<br><br><h2>Todays work:</h2><br><b>Mutagenesis</b><br>338.pSB1C3-meffBlue <br><br><b>Digest:</b><br>200.pSB1C3-B0034-asCP E,X<br>148.pSB1C3-RBS-gfasPurple E,X<br>20.pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1 E,X<br>257. pSB1C3-pemp (two tubes) with Pem-mutE_F & Pem-mutE_R (EcoRI)<br>288. pSB1C3-pem (two tubes) with Pem-mutE_F & Pem-mutE_R (EcoRI)<br><br><b>Ligation:</b><br>200.pSB1C3-B0034-asCP+91.BBa_J61002 (amp)-J23106→<br>→ 343.J23106-B0034-asCP<br>148.pSB1C3-RBS-gfasPurple+91.BBa_J61002 (amp)-J23106→<br>→ 160.j23106-RBS-gfasPurple<br>148.pSB1C3-RBS-gfasPurple+92.BBa_J61002 (amp)-J23110→<br>→ 159j23110-RBS-gfasPurple<br>20.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1+6.pSB1K3-red→<br>→215.pSB1K3-pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>184.pSB1C3-B0034-CrtZ+6.pSB1K3-Red+103.pSB1K3-J23101→<br>→ 303.pSB1K3-J23101-B0034-CrtZ<br>127.pSB1A3-B0034-UGTCs2-His+7.pSB3K3-Red+CP8→<br>→ 223.pSB3K3-CP8-B0034-UGTCs2-His<br><br><b>Transformation:</b><br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>208.pSB1A3-B0034-Thioredoxin-ZCD-His<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>343.pSB1C3-J23106-B0034-asCP<br>160.pSB1C3-j23106-RBS-gfasPurple<br>159.pSB1C3-j23110-RBS-gfasPurple<br>303.pSB1K3-j23101-B0034-CrtZ<br>269.pSB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>336.pSB1C3.spisPink<br>337.pSB1C3.scOrange<br>284.pSB1C3.B0034-aeBlue<br>285.pSB1C3.J23110-B0034-aeBlue<br>286.pSB1C3.J23110-B0034-amilCP<br>287.pSB1C3-J23110-B0034-asCP<br>336.pSB1C3-spisPink<br>337.pSB1C3-scOrange<br>284. pSB1C3-B0034-aeBlue<br>285. pSB1C3-J23110-B0034-aeBlue<br>286. pSB1C3-J23110-B0034-amilCP<br>287. pSB1C3-J23110-B0034-asCP<br><br><b>O/N screening:</b><br>339. pSB1C3-gfasPurple<br>340. pSB1C3-amajCP(amajLime)<br>341. pSB1C3-tsPurple<br>342. pSB1C3-fwYellow<br>Lb31. plantarum 36E pVS2<br>Lb32. plantarum 36E pSBLB4B15-CP30-fusionGFP<br>Lb33. plantarum 36E pSBLB4B15-CP11-usp45-fusionGFP<br>Lb34. plantarum 36E pSBLB4B15-CP11-tsPurple<br>Lb35. reuteri DSM 20016 pVS2<br>Lb36. reuteri DSM 20016 pSBLB4B15-CP30-mCherry<br>Lb37. reuteri DSM 20016 pSBLB4B15-CP30-amilGFP<br>Lb38. reuteri DSM 20016 pSBLB4B15-CP11-amilCP<br>Lb39. reuteri DSM 20016 pSBLB4B15-CP30-amilCP<br>Lb40. reuteri DSM 20016 pSBLB4C15-CP11-aeBlue<br>Lb41. reuteri DSM 20016 pSBLB4C15-CP30-aeBlue<br>Lb42. reuteri DSM 20016 pSBLB4C15-CP1-BFP<br>Lb43. reuteri DSM 20016 pSBLB4C15-CP8-BFP<br>Lb44. reuteri DSM 20016 pSBLB4C15-CP11-BFP<br>Lb45. reuteri DSM 20016 pSBLB4C15-CP29-BFP<br>Lb46. reuteri DSM 20016 pSBLB4C15-CP30-BFP<br>136.<br>312<br>313<br>315<br>321<br>325<br>327<br>332<br>317<br><br><b>O/N:</b><br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>143. pEL3S17-CP8-B0034-UGTCs2-6xHis<br>179. pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180. pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>265. J23110-B0034-Exp-Linker-Miraculin-His<br>301. pEL3S17-J23110-B0034-Thioredoxin<br><br><b>Plasmid prepp & sequence prepp</b><br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br><br><b>Gel electrophoresis:</b><br>Lb31. plantarum 36E pVS2<br>Lb32. plantarum 36E pSBLB4B15-CP30-fusionGFP<br>Lb33. plantarum 36E pSBLB4B15-CP11-usp45-fusionGFP<br>Lb34. plantarum 36E pSBLB4B15-CP11-tsPurple<br>Lb35. reuteri DSM 20016 pVS2<br>Lb36. reuteri DSM 20016 pSBLB4B15-CP30-mCherry<br>Lb37. reuteri DSM 20016 pSBLB4B15-CP30-amilGFP<br>Lb38. reuteri DSM 20016 pSBLB4B15-CP11-amilCP<br>Lb39. reuteri DSM 20016 pSBLB4B15-CP30-amilCP<br>Lb40. reuteri DSM 20016 pSBLB4C15-CP11-aeBlue<br>Lb41. reuteri DSM 20016 pSBLB4C15-CP30-aeBlue<br>Lb42. reuteri DSM 20016 pSBLB4C15-CP1-BFP<br>Lb43. reuteri DSM 20016 pSBLB4C15-CP8-BFP<br>Lb44. reuteri DSM 20016 pSBLB4C15-CP11-BFP<br>Lb45. reuteri DSM 20016 pSBLB4C15-CP29-BFP<br>Lb46. reuteri DSM 20016 pSBLB4C15-CP30-BFP<br>257. pSB1C3-pemp (two tubes) with Pem-mutE_F & Pem-mutE_R<br>288. pSB1C3-pem (two tubes) with Pem-mutE_F & Pem-mutE_R<br><i>+ E. coli transformations</i><br><br><b>PCR-purification:</b><br>257. pSB1C3-pemp (two tubes) with Pem-mutE_F & Pem-mutE_R<br>288. pSB1C3-pem (two tubes) with Pem-mutE_F & Pem-mutE_R<br><br><b>DpnI:</b><br>257. pSB1C3-pemp (two tubes) with Pem-mutE_F & Pem-mutE_R<br>288. pSB1C3-pem (two tubes) with Pem-mutE_F & Pem-mutE_R<br><br><h2>Results:</h2><br><b>Transformations:</b><br>All transformations spread on Bleomycin/zeocin have grown! If they are the right lactobacilli with the right plasmid remains to be seen, there seem to be a lot of contamination in the form of “clouds” covering all plates. Yet still there are larger colonies that very well could be what we want. Maybe the antibiotics are not efficient enough, we will try to spread normal plasmid free reuteri from frozen stock on the plates to investigate… We’ll also try a colony screening today with VF2 and VR. <br>Nothing grew on chloramphenicol plates, not even the positive control pVS2, though it grew on erythromycin both when transformed into reuteri and plantarum. <br><br><b>Gel electrophoresis:</b><br>All Pem/Pemp mutagenesises showed bands at the right lengths (~2700 bp), although they were a bit smudgy - weakly stretching backward toward the wells. The digests of Pem simply showed one band, a weaker counterpart of the uncut PCR-product. We suspect the digest didn’t go as supposed, maybe due to the PCR buffer. <br>The lactobacillus colony screenings showed up empty, perhaps the cells didn’t lyse? Or didn’t contain plasmids with VF and VR… We will continue to investigate the resistance problems, and await the sequencing results of the insertsite.<br>The screenings of e. coli transformations done thursday also gave us an empty gel, without even ladders. Possibly the gel had to shallow wells.</div>' | ||
| + | } | ||
| + | else if(id == 'd2013825') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Sunday 2013-08-25</h1><br><b>Name of participants:</b> Alexander, Pontus, Sabri, Malin, Marcus H, Karl H, Hampus.E, Lovisa P, Emil M<br><br><h2>Ongoing constructs:</h2><br>102.pSB1C3-CrtE<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>(201. E.Coli. Nissle - CrtY-CrtI)<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br>305.pSB1K3-J23110-CrtZ<br>308.pSB1C3-Miraculin<br>309.pSB1C3-B0034-Miraculin<br>261. pSB3K3-J23110-B0034-His-4CL<br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br><br><h2>Todays work:</h2><br><b>Mutagenesis</b><br>338.pSB1C3-meffBlue<br><br><b>Plasmidprepp:</b><br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>265.pSB3K3.J23110-B0034-Exp-Linker-Miraculin-His<br>339. pSB1C3-gfasPurple<br>340. pSB1C3-amajCP(amajLime)<br>341. pSB1C3-tsPurple<br>342. pSB1C3-fwYellow<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>143. pEL3S17-CP8-B0034-UGTCs2-6xHis<br>179. pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180. pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>265. J23110-B0034-Exp-Linker-Miraculin-His<br>301. pEL3S17-J23110-B0034-Thioredoxin<br><br><b>Digest:</b><br>147.pSB1C3-B0034-Thioredoxin<br><br><b>Ligate:</b><br>147.pSB1C3-B0034-Thioredoxin+CP1→<br>→ 346.pSB1C3-CP1-B0034-Thioredoxin<br>147.pSB1C3-B0034-Thioredoxin+CP8→<br>→347.pSB1C3-CP8-B0034-Thioredoxin<br>147.pSB1C3-B0034-Thioredoxin+CP41→<br>→348.pSB1C3-CP41-B0034-Thioredoxin<br><br><b>Transformation:</b><br>1.pSB1C3-Red (competens test)<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>208.pSB1A3-B0034-Thioredoxin-ZCD-His<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>269.pSB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>285.pSB1C3.J23110-B0034-aeBlue<br>286.pSB1C3.J23110-B0034-amilCP<br>287.pSB1C3-J23110-B0034-asCP303.pSB1K3-j23101-B0034-CrtZ<br>336.pSB1C3-spisPink<br>337pSB1C3-scOrange<br>346.pSB1C3-CP1-B0034-Thioredoxin<br>347.pSB1C3-CP8-B0034-Thioredoxin<br>348.pSB1C3-CP41-B0034-Thioredoxin<br>285. pSB1C3-J23110-B0034-aeBlue<br>286. pSB1C3-J23110-B0034-amilCP<br>287. pSB1C3-J23110-B0034-asCP<br>336.pSB1C3-spisPink<br>337.pSB1C3-scOrange<br>159. pSB1C3-J23110-RBS-gfasPurple<br>160. pSB1C3-J23110-RBS-gfasPurple<br><br><b>Re-streak:</b><br>284. pSB1C3-B0034-aeBlue<br><br><b>Sequenceprep:</b><br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br>339. pSB1C3-gfasPurple<br>340. pSB1C3-amajCP(amajLime)<br>341. pSB1C3-tsPurple<br>342. pSB1C3-fwYellow<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013826') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-08-26</h1><br><b>Name of participants: </b>Alona Nyberg, Mikael Strandgren, Anders Edlund, Marcus H, Karl H, Hampus.E, Lovisa P, Emil M<br><br><h2>Ongoing constructs:</h2><br>261. pSB3K3-J23110-B0034-His-4CL<br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br>349. pSB3K3-B0034-TAL(mut)-B0034-4CL-B0034-STS<br>102.pSB1C3-CrtE<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>(201. E.Coli. Nissle - CrtY-CrtI)<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br>305.pSB1K3-J23110-CrtZ<br>308.pSB1C3-Miraculin<br>309.pSB1C3-B0034-Miraculin<br><br><h2>Todays work</h2><br><b>Streak from frozen stock:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>139. pSB3K3-J23101-B0034-BFP<br><br><b>PCR:</b><br>Lb9. faecalis JH2-2 pAMbeta1-ori with pAM-d-BII_F and pAM-s-X1_R<br>77. pSB4C15-red with pEL-C1_F and pEL-d-BII_R<br>77. pSB4C15-red with pEL-C1_F and pEL-d-BII_R*<br><i>*(with annealing temperatures +2° and +4°C)</i><br>Slovenian 4CL with His-primers<br><br><b>Gel electrophoresis:</b><br>Erythromycin-cassette PCR-product from Lb2.1 pLUL631<br>Erythromycin-cassette PCR-product from Lb2.2 pLUL631<br>Lb9. faecalis JH2-2 pAMbeta1-ori with pAM-d-BII_F and pAM-s-X1_R<br>77. pSB4C15-red with pEL-C1_F and pEL-d-BII_R<br><br><b>Assembly & transformation:</b><br>349<br><br><b>Mutagenesis</b><br>338.pSB1C3-meffBlue<br><br><b>Re streak</b><br>285. pSB1C3-J23110-B0034-aeBlue<br>286. pSB1C3-J23110-B0034-amilCP<br>287. pSB1C3-J23110-B0034-asCP<br>336.pSB1C3-spisPink<br>337.pSB1C3-scOrange<br>159. pSB1C3-J23110-RBS-gfasPurple<br>160. pSB1C3-J23110-RBS-gfasPurple<br>343. pSB1C3-J23106-B0034-asCP<br><br><b>Plasmidprepp:</b><br>102.pSB1C3-CrtE<br>272.pSB1K3-B0034-Thioredoxin-ZCD-His<br>(272F.pSB1K3-B0034-Thioredoxin-ZCD-His)<br>308.pSB1C3-Miraculin<br>309.pSB1C3-B0034-Miraculin<br><br><b>O/N screen</b><br>284. pSB1C3-B0034-aeBlue<br><br><b>Screening:</b><br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>269.pSB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>303.pSB1K3-j23101-B0034-CrtZ<br>346.pSB1C3-CP1-B0034-Thioredoxin<br>347.pSB1C3-CP8-B0034-Thioredoxin<br>348. pSB1C3-CP41-B0034-Thioredoxin<br><br><b>O/N:</b><br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>269.pSB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>303.pSB1K3-j23101-B0034-CrtZ<br>269.pSB3K3-CP8-B0034-Exp-Linker-Miraculin-His (annan ligering än ovan, den som screenades)<br><br><b>Re-streak</b><br>269.pSB3K3-CP8-B0034-Exp-Linker-Miraculin-His (den som screenades)<br><br><b>Sekvensprep:</b><br>102.pSB1C3-CrtE<br>272.pSB1K3-B0034-Thioredoxin-ZCD-His<br>(272F.pSB1K3-B0034-Thioredoxin-ZCD-His)<br>308.pSB1C3-Miraculin<br>309.pSB1C3-B0034-Miraculin<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>143. pEL3S17-CP8-B0034-UGTCs2-6xHis<br>179. pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>265. J23110-B0034-Exp-Linker-Miraculin-His<br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>339. pSB1C3-gfasPurple<br>340. pSB1C3-amajCP(amajLime)<br>341. pSB1C3-tsPurple<br>342. pSB1C3-fwYellow<br><br><b>Ligation:</b><br>6. pSB1K3-Red + 147.pSB1C3-B0034-Thioredoxin +CP1→<br>→ 346.pSB1K3-CP1-B0034-Thioredoxin<br>6. pSB1K3-Red + 147.pSB1C3-B0034-Thioredoxin +CP8→<br>→ 346.pSB1K3-CP1-B0034-Thioredoxin<br>6. pSB1K3-Red + 147.pSB1C3-B0034-Thioredoxin +CP41→<br>→ 346.pSB1K3-CP1-B0034-Thioredoxin<br>6. pSB1K3-Red + 20.1.pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1 → <br>→215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>184.pSB1C3-B0034-CrtZ+6.pSB1K3-Red+103.pSB1K3-J23101→<br>→ 303.pSB1K3-J23101-B0034-CrtZ<br>127.pSB1A3-B0034-UGTCs2-His+7.pSB3K3-Red+CP8→<br>→ 223.pSB3K3-CP8-B0034-UGTCs2-His<br><br><h2>Results:</h2><br><b>Gel electrophoresis: </b><br>Erythromycin-cassette PCR-product: Good bands! → proceed with PCR-cleanup and assembly<br>Erythromycin-cassette PCR-product: --”--<br>Lb9. faecalis JH2-2 pAMbeta1-ori: One of the two had a good band → proceed<br>77. pSB4C15-red: Unspecific binding → redo PCR with +2° and +4° annealing temp.<br>269 showed correct bands. 247 and 248 showed credible bands, but a mistake in the assembly was detected (resistance on source of insert is the same as target backbone). These samples were discarded and a new ligation with the backbone pSB1K3 was made.<br><br><h2>Characterization:</h2><br><b>Western blot sample preparation:</b><br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br><br><h2>Other experiments</h2><br><b>Preparing MRS-agar</b><br><br><b>Preparing low salt LB-agar</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013827') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-08-27</h1><br><b>Name of participants: </b>Viktor Törnblom, Viktor Blomkvist, Alona Nyberg, Mikael Strandgren, Marcus H, Karl H, Hampus.E, Lovisa P, Emil M, Alexander, Pontus, Sabri, Magnus<br><br><h2>Ongoing constructs:</h2><br>102.pSB1C3-CrtE<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>(201. E.Coli. Nissle - CrtY-CrtI)<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br>305.pSB1K3-J23110-CrtZ<br>308.pSB1C3-Miraculin<br>309.pSB1C3-B0034-Miraculin<br>377.pSB1A3-CP8-B0034-UGTCs2-His<br>378.pSB1A3-J23110-B0034-UGTCs2-His<br>379.pSB1K3-CP8-B0034-Thioredoxin-ZCD-His<br>380.pSB1K3-J23110-B0034-Thioredoxin-ZCD-His<br>381.pSB1C3-CP8-B0034-Exp-Linker-Miraculin-Linker-His<br>382.pSB1C3-J23110-B0034-Exp-Linker-Miraculin-Linker-His<br>261. pSB3K3-J23110-B0034-His-4CL<br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br>349. pSB3K3-B0034-TAL(mut)-B0034-4CL-B0034-STS<br><br><h2>Todays work</h2><br><b>O/N:</b><br>351.1 pSB1C3-pemp-mut1<br>351.2 pSB1C3-pemp-mut1<br><br><b>Transformation:</b><br>Chromoproteins into pSB3K3<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>303.pSB1K3-j23101-B0034-CrtZ<br>346.pSB1K3-CP1-B0034-Thioredoxin<br>347.pSB1K3-CP8-B0034-Thioredoxin<br>348.pSB1K3-CP41-B0034-Thioredoxin<br><br><b>Gel electrophoresis:</b><br>77. pSB4C15-red with pEL-C1_F and pEL-d-BII_R*<br><i>*(with annealing temperatures +2° and +4°C)</i><br><br><b>Screening PCR: </b><br>351.1 pSB1C3-pemp-mut1<br>351.2 pSB1C3-pemp-mut1<br>377.pSB1A3-CP8-B0034-UGTCs2-His<br>378.pSB1A3-J23110-B0034-UGTCs2-His<br>379.pSB1K3-CP8-B0034-Thioredoxin-ZCD-His<br>380.pSB1K3-J23110-B0034-Thioredoxin-ZCD-His<br>381.pSB1C3-CP8-B0034-Exp-Linker-Miraculin-Linker-His<br>382.pSB1C3-J23110-B0034-Exp-Linker-Miraculin-Linker-His<br><br><b>Mutagenesis</b><br>338.pSB1C3-meffBlue<br><br><b>Plasmidprepp:</b><br>182.pSBLb4C5-CP8-B0034-Exp-Linker-Miraculin-His<br>269.pSB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>303.pSB1K3-j23101-B0034-CrtZ<br><br><b>Sekvensprep:</b><br>182.pSBLb4C5-CP8-B0034-Exp-Linker-Miraculin-His<br>269.pSB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>303.pSB1K3-j23101-B0034-CrtZ<br>284. pSB1C3-B0034-aeBlue<br><br><b>Re-streak</b><br>285. pSB1C3-J23110-B0034-aeBlue<br>286. pSB1C3-J23110-B0034-amilCP<br>287. pSB1C3-J23110-B0034-asCP<br>336.pSB1C3-spisPink<br>337.pSB1C3-scOrange<br>159. pSB1C3-J23110-RBS-gfasPurple<br>160. pSB1C3-J23110-RBS-gfasPurple<br>343. pSB1C3-J23106-B0034-asCP<br><br><b>PCR:</b><br>77. pSB4C15-red with pEL-C1_F and pEL-d-BII_R*<br><i>*(with annealing temperatures Tm1 and +5°C (with touchdown))</i><br><br><b>Streaking of plasmid free stock on test plates:</b><br><i>- e. coli (10. and 60.3) onto erythromycin LB-plates:</i><br>5 µg/ml<br>10 µg/ml<br>20 µg/ml<br>50 µg/ml<br><i>- lactobacillus (Lb4., Lb11. and Lb12.) onto Zeocin (bleomycin) MRS-plates of varying concentrations:</i><br>50 µg/ml<br>100 µg/ml<br><br><h2>Results</h2><br><b>Gel electrophoresis:</b><br>77. pSB4C15-red +2°C annealing temperature: Strange band at 1000 bp<br>77. pSB4C15-red +4°C annealing temperature: Strange band at 1000 bp<br><br><h2>Other experiments</h2><br><b>Preparing test plates, to determine minimum antibiotic concentration:</b><br>Lb-erythromycin:<br>5 µg/ml<br>10 µg/ml<br>20 µg/ml<br>50 µg/ml<br><br><b>MRS-zeocin:</b><br>50 µg/ml<br>100 µg/ml</div>' | ||
| + | } | ||
| + | else if(id == 'd2013828') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-08-28</h1><br><b>Name of participants: </b>Viktor Törnblom, Viktor Blomkvist, Alona Nyberg, Mikael Strandgren, Anders Edlund, Marcus H, Karl H, Hampus.E, Lovisa P, Emil M, Alexander, Pontus, Sabri<br><br><h2>Ongoing constructs:</h2><br>261. pSB3K3-J23110-B0034-His-4CL<br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br>349. pSB3K3-B0034-TAL(mut)-B0034-4CL-B0034-STS<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>282.pSB3K3-J23110-B0034-amilCP<br>283.pSB3K3-J23110-B0034-asCCP<br>299.pEL3S17-J23101-B0034-Thioredoxin<br>300.pEL3S17-J23106-B0034-Thioredoxin<br>301.pEL3S17-J23110-B0034-Thioredoxin<br><br><h2>Todays work</h2><br><b>Gel electrophoresis: </b><br>351.1 pSB1C3-pemp-mut1<br>351.2 pSB1C3-pemp-mut1<br>77. pSB4C15-red (annealing at Tm1): <br>77. pSB4C15-red (annealing at Tm1+5): <br><br><b>Ligation: </b><br>352. pSBLbC3-red (pEL4C15 (ClaI) + pVS2 (ClaI))<br>108.pEL3S17-J23106-B0034-IspA<br>140.pEL3S17-J23110-B0034-UGTCs2-His<br>141.pEL3S17-J23101-B0034-UGTCs2-His<br>142.pEL3S17-CP1-B0034-UGTCs2-His<br>143.pEL3S17-CP8-B0034-UGTCs2-His<br><br><b>PCR: </b><br>77. pSB4C15-red with pEL-C1_F and pEL-d-BII_R (Tm1)<br><br><b>Restreak</b><br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br><br><b>Screening O/N:</b><br>109.pEL3S17-K206000-B0034-IspA <br>110.pEL3S17-K206001-B0034-IspA <br>147.B0034-Thioredoxin<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His <br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His <br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His <br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His <br>267.psB3K3-K206000-B0034-IspA <br>268.psB3K3-K206001-B0034-IspA <br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His <br>300.J23106-B0034-Thioredoxin<br>202.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br><br><br><b>Digest</b><br>36. pEL3S17-red<br><br><b>Subclone:</b><br>280. pSB1A3-B0034-aeBlue à 284. pSB1C3-B0034-aeBlue<br>281. pSB1K3-J23110-B0034-aeBlue à 285. pSB1C3-J23110-B0034-aeBlue<br>282. pSB3K3-J23110-B0034-amilCP à 286. pSB1C3-J23110-B0034-amilCP<br>283. pSB3K3-J23110-B0034-asCP à 287. pSB1C3-J23110-B0034-asCP<br><br><br><b>Two step Assembly:</b><br>92. BBa_J61002-J23110, 149. pSB1C3-RBS-meffRed à 155. pSB1C3-J23110-RBS-meffRed<br>91. BBa_J61002-J23106, 149. pSB1C3-RBS-meffRed à 156. pSB1C3-J23106-RBS-meffRed<br>92. BBa_J61002-J23110, 150. pSB1C3-RBS-spisPink à 157. pSB1C3-J23110-RBS-spisPink<br>91. BBa_J61002-J23106, 150. pSB1C3-RBS-spisPink à 158.pSB1C3-J23106-RBS-spisPink<br>92. BBa_J61002-J23110, 148. pSB1C3-RBS-gfasPurple à 159.pSB1C3-J23110-RBS-gfasPurple<br>91. BBa_J61002-J23106, 148 pSB1C3-RBS-gfasPurple à 160.pSB1C3-J23106-RBS-gfasPurple<br><br><h2>Results</h2><br><b>Streaking of plasmid free stock on test plates:</b><br>- e. coli (10. and 60.3) onto erythromycin LB-plates:<br>5 µg/ml: Growth <br>10 µg/ml: Growth<br>20 µg/ml: Growth<br>50 µg/ml: Still some growth, but less → do new plates with higher conc. and try 50 µg/ml-plates from Erik Gullberg in case our antibiotic has lost efficiency.<br>- lactobacillus (Lb4., Lb11. and Lb12.) onto Zeocin (bleomycin) MRS-plates of varying concentrations:<br>50 µg/ml: Growth<br>100 µg/ml: Growth (Plantarum grew less, but did so also on antibiotic free plates.)<br>- lactobacillus (Lb4., Lb11. and Lb12.) onto antibiotic-free MRS-plates:<br>Everything grew on Ab-, plantarum slightly less.<br><br><b>Gel electrophoresis: </b><br>351.1 pSB1C3-pemp-mut1: A weak band at ~1000 bp<br>351.2 pSB1C3-pemp-mut1: Nothing<br>77. pSB4C15-red (annealing at Tm1): Nothing → do a new PCR at TM1.<br>77. pSB4C15-red (annealing at Tm1+5): Nothing. <br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268: Failed<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1: Failed<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1: Failed<br><br><b>Re-streak:</b><br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br><br><h2>Other experiments</h2><br><b>Preparation of new test-plates:</b><br>LB-erythromycin:<br>100 µg/ml<br>200 µg/ml</div>' | ||
| + | } | ||
| + | else if(id == 'd2013829') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-08-29</h1><br><b>Name of participants: </b>Mikael Strandgren, Viktor Blomkvist, Alona Nyberg, Anders Edlund, Marcus H, Karl H, Hampus.E, Lovisa P, Emil M, Alexander, Pontus, Sabri, Magnus<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>353. pSBLb4E15<br>261. pSB3K3-J23110-B0034-His-4CL<br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br>349. pSB3K3-B0034-TAL(mut)-B0034-4CL-B0034-STS<br>19.pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>20.pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>102.pSB1C3-CrtE<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>(201. E.Coli. Nissle - CrtY-CrtI)<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br>305.pSB1K3-J23110-CrtZ<br>308.pSB1C3-Miraculin<br>309.pSB1C3-B0034-Miraculin<br>377.pSB1A3-CP8-B0034-UGTCs2-His<br>378.pSB1A3-J23110-B0034-UGTCs2-His<br>379.pSB1K3-CP8-B0034-Thioredoxin-ZCD-His<br>380.pSB1K3-J23110-B0034-Thioredoxin-ZCD-His<br>381.pSB1C3-CP8-B0034-Exp-Linker-Miraculin-Linker-His<br>382.pSB1C3-J23110-B0034-Exp-Linker-Miraculin-Linker-His<br><br><h2>Todays work</h2><br><b>Transformation:</b><br>353. pSBLb4E15<br><br><b>Plasmidprep:</b><br>223<br>346<br><br><h2>Results</h2><br><b>Streaking of plasmid free stock on test plates:</b><br>- e. coli (10. and 60.3) onto erythromycin LB-plates:<br>100 µg/ml: Weak growth<br>200 µg/ml: Possibly no growth, let them “grow” some more. <br><br><h2>Characterization experiments:</h2><br><b>Western blot on construct:</b><br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br><br><h2>Other experiments</h2><br><b>Preparation of new test-plates:</b><br>MRS-Zeocin:<br>250 µg/ml<br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013830') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-08-30</h1><br><b>Name of participants: </b>Mikael Strandgren, Viktor Björklund, Alona Nyberg, Marcus H, Karl H, Hampus.E, Lovisa P, Emil M, Alexander, Pontus, Sabri, Magnus<br><br><h2>Ongoing constructs:</h2><br>77. pSB4C15-red <br>261. pSB3K3-J23110-B0034-His-4CL<br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br>349. pSB3K3-B0034-TAL(mut)-B0034-4CL-B0034-STS<br>19.pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>20.pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>102.pSB1C3-CrtE<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>140.pEL3S17-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>(201. E.Coli. Nissle - CrtY-CrtI)<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>272. pSB1K3-B0034-Thioredoxin-ZCD-His<br>299. pEL3S17-J23101-B0034-Thioredoxin<br>300. pEL3S17-J23106-B0034-Thioredoxin<br>301. pEL3S17-J23110-B0034-Thioredoxin<br>303. pSB1A3-j23101-B0034-CrtZ<br>304. pSB1A3-j23106-B0034-CrtZ<br>305.pSB1K3-J23110-CrtZ<br>308.pSB1C3-Miraculin<br>309.pSB1C3-B0034-Miraculin<br>377.pSB1A3-CP8-B0034-UGTCs2-His<br>378.pSB1A3-J23110-B0034-UGTCs2-His<br>379.pSB1K3-CP8-B0034-Thioredoxin-ZCD-His<br>380.pSB1K3-J23110-B0034-Thioredoxin-ZCD-His<br>381.pSB1C3-CP8-B0034-Exp-Linker-Miraculin-Linker-His<br>382.pSB1C3-J23110-B0034-Exp-Linker-Miraculin-Linker-His<br>Lb9. faecalis JH2-2 pAMbeta<br><br><h2>Todays work</h2><br><b>Gel extraction:</b><br>77. pSB4C15-red (annealing at Tm1): <br><br><b>PCR:</b><br>Lb9. faecalis JH2-2 pAMbeta1-ori with pAM-d-BII_F and pAM-s-X1_R*<br>*Since the product from monday disappeared.<br><br><b>PCR purification:</b><br>Lb9. faecalis JH2-2 pAMbeta1-ori<br><br><b>Digestion:</b><br>Lb9. faecalis JH2-2 pAMbeta1-ori (PCR product)(XbaI and BglII)<br>77. pSB4C15-red (PCR product) (NheI and BglII) <br><br><b>Ligation:</b><br>169. pSBLbEcC2-red (shuttle-vector with pAMbeta and e. coli replicon)<br><br><h2>Results</h2><br><b>Western blot:</b> <br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br>failed.<br><br><h2>Characterization experiments:</h2><br><b>Western blot on construct:</b><br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS</div>' | ||
| + | } | ||
| + | else if(id == 'd2013831') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-08-31</h1><br><b>Name of participants: </b>Mikael Strandgren, Marcus H, Karl H, Hampus.E, Lovisa P, Emil M<br><br><h2>Ongoing constructs:</h2><br>261. pSB3K3-J23110-B0034-His-4CL<br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br>349. pSB3K3-B0034-TAL(mut)-B0034-4CL-B0034-STS<br><br><h2>Todays work</h2><br><b>Transformation:</b><br>111. pSBLb4C15-red<br>169. pSBLbEcC2-red (pEL+replicon from pAMbeta1)<br>386. pSBLbEcC2-red (in PK401 dam-) (pEL+replicon from pAMbeta1)<br><br><b>Gel electrophoresis:</b><br>169. pSBLbEcC2-red (ligation)<br><br><b>Ligation:</b><br>111. pSBLb4C15-red (pEL+ replicon from pJP)<br>352. pSBLbC3-red (pEL+ replicon from pVS2)<br><br><h2>Results</h2><br><b>Gel electrophoresis:</b><br>169. pSBLbEcC2-red (ligation): No band, possibly to weak due to being ligation → transform and screen.</div>' | ||
| + | } | ||
| + | else if(id == 'd201391') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Sunday 2013-09-01</h1><br><br><b>Name of participants: </b>Anders Edlund, Mikael Strandgren, Alona Nyberg, Marcus H, Karl H, Hampus.E, Lovisa P, Emil M<br><br><h2>Ongoing constructs:</h2><br>111. pSBLb4C15-red<br>169. pSBLbEcC2-red<br>275. pSBLb4C15-red (dam-)<br>352. pSBLbC3-red<br>406. pSBLbC3-red (dam-)<br>261. pSB3K3-J23110-B0034-His-4CL<br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br>349. pSB3K3-B0034-TAL(mut)-B0034-4CL-B0034-STS<br><br><h2>Todays work</h2><br><b>Transformation:</b><br>111. pSBLb4C15-red (pEL+ replicon from pJP)<br>275. pSBLb4C15-red (dam-)<br>352. pSBLbC3-red (pEL+ replicon from pVS2)<br>406. pSBLbC3-red (dam-)<br><br><b>Screening:</b><br>169. pSBLbEcC2-red (pEL+replicon from pAMbeta1) (x2)<br><br><b>Re-streak:</b><br>169. pSBLbEcC2-red (pEL+replicon from pAMbeta1)<br><br><h2>Results</h2><br>First screening failed (weird gel)<br><br><h2>Characterization experiments:</h2><br><b>Western blot on construct:</b><br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS</div>' | ||
| + | } | ||
| + | else if(id == 'd201392') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-09-02</h1><br><br><b>Name of participants:</b> Anders Edlund, Mikael Strandgren, Marcus H, Karl H, Hampus.E, Lovisa P, Emil M<br><br><h2>Ongoing constructs: </h2><br>111. pSBLb4C15-red<br>169. pSBLbEcC2-red<br>352. pSBLbC3-red<br>261. pSB3K3-J23110-B0034-His-4CL<br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br>349. pSB3K3-B0034-TAL(mut)-B0034-4CL-B0034-STS<br><br><h2>Todays work: </h2><br><b>O/N:</b><br>169. pSBLbEcC2-red<br><br><b>Mutagenesis PCR:</b><br>111.7 pSBLb4C15-red *<br>*with primers to change the in the chloramphenicol-cassette promotor into one from the CP-serie. (CP11, CP29 and CP30 were tried). Two attempts, second over night.<br><br><b>Ligation:</b><br>111. pSBLb4C15-red (pEL4C15+replicon from pJP059)*<br>352. pSBLbC3-red*<br>*New attempts at ligation, just in case.<br><br><h2>Results: </h2><br><b>Gel electrophoresis:</b><br>111.7 pSBLb4C15-red: No bands<br><br><b>Western blot on construct:</b><br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS<br>failed.<br><br><h2>Characterization experiments:</h2><br><b>Western blot on construct:</b><br>333. pSB3K3-J23110-HIS-STS<br>334. pSB3K3-CP1-His-STS<br>335. pSB3K3-J23101-HIs-STS</div>' | ||
| + | } | ||
| + | else if(id == 'd201393') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-09-03</h1><br><br><b>Name of participants: </b>Anders Edlund, Mikael Strandgren, Marcus H, Karl H, Hampus.E, Lovisa P, Emil M<br><br><h2>Ongoing constructs: </h2><br>111. pSBLb4C15-red<br>169. pSBLbEcC2-red<br>352. pSBLbC3-red<br>261. pSB3K3-J23110-B0034-His-4CL<br>335. pSB3K3-J23101-HIs-STS<br>349. pSB3K3-B0034-TAL(mut)-B0034-4CL-B0034-STS<br><br><h2>Todays work: </h2><br>Frozen stock and plasmid preparation:<br>169. pSBLbEcC2-red<br><br><b>Gel electrophoresis:</b><br>111.7 pSBLb4C15-red* <br><i>*to change the in the chloramphenicol-cassette promotor into one from the CP-serie. (CP11, CP29 and CP30 were tried)</i><br><br><b>Transformation:</b><br>111. pSBLb4C15-red (pEL4C15+replicon from pJP059)*<br><i>*New attempts at ligation, just in case.</i><br><br><b>Sequencing preparation:</b><br>169. pSBLbEcC2-red<br><br><h2>Characterization experiments:</h2><br><b>Western blot sample preparation on construct:</b><br>335. pSB3K3-J23101-HIs-STS<br><br><h2>Results: </h2><br><b>Frozen stock and plasmid preparation:</b><br>169. pSBLbEcC2-red: 44.2 ng/µl<br>169. pSBLbEcC2-red: 47.8 ng/µl<br><br><b>Gel electrophoresis:</b><br>111.7 pSBLb4C15-red: Correct bands in five lanes of six, weird ladder.</div>' | ||
| + | } | ||
| + | else if(id == 'd201394') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-09-04</h1><br><br><b>Name of participants: </b>Anders Edlund, Mikael Strandgren, Stephanie Herman, Marcus H, Karl H, Hampus.E, Lovisa P, Emil M<br><br><h2>Ongoing constructs: </h2><br>111. pSBLb4C15-red<br>169. pSBLbEcC2-red<br>352. pSBLbC3-red<br>335. pSB3K3-J23101-HIs-STS<br><br><h2>Todays work: </h2><br><b>Assembly:</b><br>353. pSBLb4E15-red (ermR-cassette from three different sources, 2.1, 2.3 and old)<br><br><b>Trasformation:</b><br>111.7 (old, used in earlier attempts of ermR, cut with SacI+SalI and ligated to test RFP)<br>353. pSBLb4E15-red <br><br><b>PCR-purification:</b><br>111.7 pSBLb4C15-red* <br>*to change the in the chloramphenicol-cassette promotor into one from the CP-serie. (CP11, CP29 and CP30 were tried)<br><br><b>DpnI-treatment:</b><br>111.7 pSBLb4C15-red* <br>*to change the in the chloramphenicol-cassette promotor into one from the CP-serie. (CP11, CP29 and CP30 were tried)</div>' | ||
| + | } | ||
| + | else if(id == 'd201395') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-09-05</h1><br><b>Name of participants:</b> Marcus H, Karl H, Hampus.E, Lovisa P, Emil M, Christoffer L<br><br><h2>Ongoing constructs:</h2><br>335. pSB3K3-J23101-HIs-STS<br><br><h2>Todays work</h2><b>First day of survival test:</b><br>O/N in antibiotic-free medium and re-streak on plates with and without antibiotics.<br><br><h2>Characterization:</h2><br><b>Western blot on construct: </b><br>335. pSB3K3-J23101-HIs-STS</div>' | ||
| + | } | ||
| + | else if(id == 'd201396') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-09-06</h1><br><br><b>Name of participants: </b>Mikael Strandgren, Marcus H, Karl H, Hampus.E, Lovisa P, Emil M, Christoffer L<br><br><h2>Ongoing constructs: </h2><br>335. pSB3K3-J23101-HIs-STS<br><br><h2>Todays work: </h2><br><b>PCR-purification and recircularization:</b><br>Mutagenesis´d pSBLb4C15 with promotor in CmR changed to CP11, CP29 or CP30)<br><br><b>O/N:</b><br>111.7 pSBLb4C15-red*<br><i>*to do a new plasmid preparation since old may have corrupt red-cassette/contamination)</i><br><br><b>Streak from frozen stock:</b><br>111.7 pSBLb4C15-red*<br><i>* to test if red-cassette has been damaged, or possibly the stock contaminated.</i><br><br><b>Second day of survival test:</b><br>O/N in antibiotic-free medium and re-streak on plates with and without antibiotics.<br><br><h2>Characterization:</h2><br><b>Western blot on construct: </b><br>335. pSB3K3-J23101-HIs-STS</div>' | ||
| + | } | ||
| + | else if(id == 'd201397') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-09-07</h1><br><br><b>Name of participants:</b> Mikael Strandgren, Christoffer L<br><br><h2>Ongoing constructs: </h2><br>407. pSB1C3-CP29-Mcherry<br>408. pSB1C3-CP30-usp45-mCherry<br>409. pSB1C3-CP29-usp45-mCherry<br>410. pSB1C3-CP30-tsPurple<br>411. pSB1C3-CP29-tsPurple <br>412. pSB1C3-CP11-tsPurple <br>413. pSB1C3-CP30-amilCP <br>414. pSB1C3-CP29-amilCP <br>415. pSB1C3-CP11-amilCP <br>416. pSB1C3-CP30-aeBlue<br>417. pSB1C3-CP29-aeBlue<br>418. pSB1C3-CP11-aeBlue<br>419. pSBLbEcE-red<br><br><h2>Todays work:</h2> <br><b>Plasmid preparation:</b><br>351.5 pSB1C3-pemp-mut1 (mutE) <br>355.2 pSB3K3-CP29-mCherry<br>355.3 pSB3K3-CP29-mCherry<br>357.3 pSB3K3-CP30-usp45-mCherry<br>357.4 pSB3K3-CP30-usp45-mCherry<br>358.1 pSB3K3-CP29-usp45-mCherry<br>358.2 pSB3K3-CP29-usp45-mCherry<br>363. pSB3K3-CP30-tspurple<br>364. pSB3K3-CP29-tspurple<br>365. pSB3K3-CP11-tspurple<br>366. pSB3K3-CP30-amilCP<br>367. pSB3K3-CP29-amilCP<br>368. pSB3K3- CP11-amilCP<br>369. pSB3K3-CP30-aeblue<br>370. pSB3K3-CP29-aeblue<br>371. pSB3K3-CP11-aeblue<br><br><b>Assembly:</b><br>407. pSB1C3-CP29-Mcherry*<br>408. pSB1C3-CP30-usp45-mCherry*<br>409. pSB1C3-CP29-usp45-mCherry*<br>410. pSB1C3-CP30-tsPurple*<br>411. pSB1C3-CP29-tsPurple <br>412. pSB1C3-CP11-tsPurple <br>413. pSB1C3-CP30-amilCP* <br>414. pSB1C3-CP29-amilCP <br>415. pSB1C3-CP11-amilCP <br>416. pSB1C3-CP30-aeBlue*<br>417. pSB1C3-CP29-aeBlue<br>418. pSB1C3-CP11-aeBlue<br>419. pSBLbEcE-red (two variants, from two different sources of ermR. 419.1=2.3, 419.2=old)<br>* pSB1C3 ran out after six ligations. New was made and ligated later, but couldn’t be included in the transformation today.<br><br><b>Transformation</b><br>411. pSB1C3-CP29-tsPurple <br>412. pSB1C3-CP11-tsPurple <br>414. pSB1C3-CP29-amilCP <br>415. pSB1C3-CP11-amilCP <br>417. pSB1C3-CP29-aeBlue <br>418. pSB1C3-CP11-aeBlue<br>419. pSBLbEcE-red <br>pSBLb4C15-red with CP11, CP29 or CP30 in cmR-cassette.<br><br><b>O/N:</b><br>111.7 pSBLb4C15-red<br><br><b>Third day of survival test:</b><br>O/N in antibiotic-free medium and re-streak on plates with and without antibiotics.<br><br><h2>Results: </h2><br><b>O/N:</b><br>111.7 pSBLb4C15-red: Little growth, since 15ml-tube had to be used, and the lid was closed to tight. → new ON today.<br><br><b>Plasmid preparation:</b><br>351.5 pSB1C3-pemp-mut1 (mutE): 205.6 ng/µl<br>355.2 pSB3K3-CP29-mCherry: 90.6 ng/µl<br>355.3 pSB3K3-CP29-mCherry: 79.3 ng/µl<br>357.3 pSB3K3-CP30-usp45-mCherry: 77.3 ng/µl<br>357.4 pSB3K3-CP30-usp45-mCherry: 95.5 ng/µl<br>358.1 pSB3K3-CP29-usp45-mCherry: 90.4 ng/µl<br>358.2 pSB3K3-CP29-usp45-mCherry: 87.1 ng/µl<br>363. pSB3K3-CP30-tspurple: 75.5 ng/µl<br>364. pSB3K3-CP29-tspurple: 43.9 ng/µl<br>365. pSB3K3-CP11-tspurple: 47.3 ng/µl<br>366. pSB3K3-CP30-amilCP: 74.5 ng/µl<br>367. pSB3K3-CP29-amilCP: 77.2 ng/µl<br>368. pSB3K3- CP11-amilCP: 34.3 ng/µl<br>369. pSB3K3-CP30-aeblue: 37.4 ng/µl<br>370. pSB3K3-CP29-aeblue: 60.3 ng/µl<br>371. pSB3K3-CP11-aeblue: 76.2 ng/µl </div>' | ||
| + | } | ||
| + | else if(id == 'd201398') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Sunday 2013-09-08</h1><br><br><b>Name of participants: </b>Mikael Strandgren, Anders Edlund<br><br><h2>Ongoing constructs: </h2><br>407. pSB1C3-CP29-Mcherry<br>408. pSB1C3-CP30-usp45-mCherry<br>409. pSB1C3-CP29-usp45-mCherry<br>410. pSB1C3-CP30-tsPurple<br>411. pSB1C3-CP29-tsPurple <br>412. pSB1C3-CP11-tsPurple <br>413. pSB1C3-CP30-amilCP <br>414. pSB1C3-CP29-amilCP <br>415. pSB1C3-CP11-amilCP <br>416. pSB1C3-CP30-aeBlue<br>417. pSB1C3-CP29-aeBlue<br>418. pSB1C3-CP11-aeBlue<br>419. pSBLbEcE-red<br><br><h2>Todays work: </h2><br><b>Transformations:</b><br>411. pSB1C3-CP29-tsPurple *<br>412. pSB1C3-CP11-tsPurple *<br>414. pSB1C3-CP29-amilCP *<br>415. pSB1C3-CP11-amilCP *<br>417. pSB1C3-CP29-aeBlue *<br>418. pSB1C3-CP11-aeBlue *<br>* The ones yesterday were by mistake spread on Kanamycin-plates, the same ones are now spread on chloramphenicol with 50x-dilutions, hoping they are still good.<br><br><b>ON:</b><br>353. pSBLB4E15-red<br><br><h2>Results: </h2><br><b>Transformation</b><br>411. pSB1C3-CP29-tsPurple: Growth, but spread on kanamycin plates by mistake (source of cp:s was pSB3K3) → Spread on C-plates today.<br>412. pSB1C3-CP11-tsPurple: “ <br>414. pSB1C3-CP29-amilCP “<br>415. pSB1C3-CP11-amilCP “<br>417. pSB1C3-CP29-aeBlue “<br>418. pSB1C3-CP11-aeBlue “<br>419. pSBLbEcE-red: Growth covering the plates, only white colonies.<br>pSBLb4C15-red with CP11, CP29 or CP30 in cmR-cassette: No growth</div>' | ||
| + | } | ||
| + | else if(id == 'd201399') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-09-09</h1><br><b>Name of participants: </b> Sabri, Christoffer L<br><br><h2>Todays Work:</h2><br><b>Digest</b><br>338.pSB1C3-meffBlue<br>339. pSB1C3-gfasPurple<br>340. pSB1C3-AmajCP(AmajLime)<br>341. pSB1C3-tsPurple<br>342. pSB1C3-fwYellow<br> <br><b>Ligation</b><br>338.pSB1C3-meffBlue<br>339. pSB1C3-gfasPurple<br>340. pSB1C3-AmajCP(AmajLime)<br>341. pSB1C3-tsPurple<br>342. pSB1C3-fwYellow<br><br><b>Fourth day of survival test:</b><br>O/N in antibiotic-free medium and re-streak on plates with and without antibiotics.<br><br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013910') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tueday 2013-09-10</h1><br><b>Name of participants:</b> Sabri, Christoffer L<br><br><h2>Todays Work:</h2><br><b>Re-streak</b><br>339. pSB1C3-gfasPurple<br>340. pSB1C3-AmajCP(AmajLime)<br>341. pSB1C3-tsPurple<br>342. pSB1C3-fwYellow<br>338.pSB1C3-meffBlue<br><br><b>Fifth day of survival test:</b><br>O/N in antibiotic-free medium and re-streak on plates with and without antibiotics.<br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013911') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-09-11</h1><br><b>Name of participants: </b>Sabri<br><br><h2>Todays Work:</h2><br><b>O/N:</b><br>339. pSB1C3-gfasPurple<br>340. pSB1C3-AmajCP(AmajLime)<br>341. pSB1C3-tsPurple<br>342. pSB1C3-fwYellow<br>338.pSB1C3-meffBlue</div>' | ||
| + | } | ||
| + | else if(id == 'd2013912') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-09-12</h1><br><b>Name of participants:</b> Sabri<br><br><h2>Todays Work:</h2><br><b>Plasmid prep:</b><br>339. pSB1C3-gfasPurple<br>340. pSB1C3-AmajCP(AmajLime)<br>341. pSB1C3-tsPurple<br>342. pSB1C3-fwYellow<br>338.pSB1C3-meffBlue<br> <br><b>Sequence prep:</b><br>339. pSB1C3-gfasPurple<br>340. pSB1C3-AmajCP(AmajLime)<br>341. pSB1C3-tsPurple<br>342. pSB1C3-fwYellow<br>338.pSB1C3-meffBlue</div>' | ||
| + | } | ||
| + | else if(id == 'd20131013') | ||
| + | { | ||
| + | ds ='<h1>European Jamboree Lyon end</h1>'; | ||
| + | } | ||
| + | else if(id == 'd20131012') | ||
| + | { | ||
| + | ds ='<h1>European Jamboree Lyon</h1>'; | ||
| + | } | ||
| + | else if(id == 'd20131011') | ||
| + | { | ||
| + | ds ='<h1>European Jamboree Lyon start</h1>'; | ||
| + | } | ||
| + | else if(id == 'd2013104') | ||
| + | { | ||
| + | ds ='<h1>WIKIFREEZE 23.59 Boston time</h1>'; | ||
| + | } | ||
| + | else if(id == 'd2013102') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-10-02</h1><br><b>Name of participants: </b>Anders Edlund<br><br><h2>Ongoing constructs</h2><br>Lb47. reuteri 100-23 pSBLbE-red<br><br><h2>Todays work</h2><br><b>Sequencing preparation:</b><br>Lb47. reuteri 100-23 pSBLbE-red<br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013101') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-10-01</h1><br><b>Name of participants: </b>Anders Edlund<br><br><h2>Ongoing constructs </h2><br>Lb47. reuteri 100-23 pSBLbE-red<br><br><h2>Todays work</h2><br><b>Screening PCR:</b><br>Lb47. reuteri 100-23 pSBLbE-red<br><br><h2>Results</h2><br><b>Transformation:</b><br><i>To L. reuteri 100-23, L. reuteri DSM 20016 and L. plantarum 256:</i><br>Lb3. pVS2: <b>Growing</b><br>Lb13. pJP059: <b>Growing</b><br>420. pSBLbE-CP11-amilCP: Not growing<br>435. pSBLbC-CP29-amilCP: Not growing<br>440. pSBLbC-red: Not growing<br>441. pSBLbE-red: <b>Growing</b><br>All negative controls were clear, and the positive controls grew</b>.<br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013928') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-09-28</h1><br><b>Name of participants:</b> Viktor Blomkvist, Mikael Strandgren, Erik Edlund<br><br><h2>Ongoing constructs</h2><br><b>E. coli:</b><br>420. pSBLbE-CP11-amilCP<br>435. pSBLbC-CP29-amilCP<br>440. pSBLbC-red<br>441. pSBLbE-red <br><br><b>L. reuteri:</b><br>Lb3. pVS2<br><br><b>Lc. lactis:</b><br>Lb13. pJP059<br><br><h2>Todays work</h2><br><b>Transformation:</b><br><i>To L. reuteri 100-23, L. reuteri DSM 20016 and L. plantarum 256:</i><br>Lb3. pVS2<br>Lb13. pJP059<br>420. pSBLbE-CP11-amilCP<br>435. pSBLbC-CP29-amilCP<br>440. pSBLbC-red<br>441. pSBLbE-red <br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013927') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-09-27</h1><br><b>Name of participants: </b>Mikael Strandgren, Anders Edlund<br><br><h2>Ongoing constructs</h2><br>Lb4. reuteri 100-23 no plasmid<br>Lb11. Plantarum 256 no plasmid<br>Lb12. Reuteri DSM20016 no plasmid<br>420. pSBLbE-CP11-amilCP<br>435. pSBLbC-CP29-amilCP<br>440. pSBLbC-red<br>441. pSBLbE-red <br><br><h2>Todays work</h2><br><b>Overnight:</b><br>Lb4. reuteri 100-23 no plasmid<br>Lb11. Plantarum 256 no plasmid<br>Lb12. Reuteri DSM20016 no plasmid<br><br><b>Streak from frozen stock (for photos):</b><br>420. pSBLbE-CP11-amilCP<br>435. pSBLbC-CP29-amilCP<br>440. pSBLbC-red<br>441. pSBLbE-red <br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013924') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-09-24</h1><br><b>Name of participants: </b>Erik Edlund, Mikael Strandgren, Viktor Blomkvist<br><br><h2>Ongoing constructs</h2><br><br><h2>Results</h2><br><b>Transformation</b>:<br>No growth. Failed negative controls and bad electroporations encourage us to try again when we have time, probably next saturday.<br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013921') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Saturday 2013-09-21</h1><br><b>Name of participants: </b>Erik Edlund, Mikael Strandgren, Viktor Blomkvist <br><br><h2>Ongoing constructs</h2><br>435. pSBLbC-CP29-amilCP<br>440. pSBLbC-red<br>441. pSBLbE-red <br><br><h2>Todays work</h2><br><b>Transformation</b><br><i>To L. reuteri DSM 20016 and L. plantarum 256:</i><br>Lb3. pVS2 (positive control)<br>435. pSBLbC-CP29-amilCP<br>440. pSBLbC-red<br>441. pSBLbE-red <br><br><h2>Other experiments</h2><br><b>Making MRS-agar plates:</b><br>Erythromycin<br>Chloramphenicol <br>Antibiotic-free<br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013920') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-09-20</h1><br><b>Name of participants: </b>Anders Edlund<br><br><h2>Ongoing constructs</h2><br>435. pSBLbC-CP29-amilCP<br><br><h2>Todays work</h2><br><b>Plasmid preparation:</b><br>435. pSBLbC-CP29-amilCP<br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013919') | ||
| + | { | ||
| + | ds ='<h1>Uppsala iGEMs Synbioday, 12.15 to 17.00</h1>'; | ||
| + | } | ||
| + | else if(id == 'd2013918') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-09-18</h1><br><b>Name of participants: </b> Viktor Blomkvist<br><br><h2>Ongoing constructs</h2><br>435. pSBLbC-CP29-amilCP<br><br><h2>Todays work</h2><br><b>O/N:</b><br>435. pSBLbC-CP29-amilCP<br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013917') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Tuesday 2013-09-17</h1><br><b>Name of participants: </b> Mikael Strandgren, Anders Edlund<br><br><h2>Ongoing constructs</h2><br>435. pSBLbC-CP29-amilCP<br><br><h2>Todays work</h2><br><b>Re-streak:</b><br>435. pSBLbC-CP29-amilCP<br></div>' | ||
| + | } | ||
| + | else if(id == 'd2013916') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Monday 2013-09-16</h1><br><b>Name of participants: </b> Mikael Strandgren, Anders Edlund<br><br><h2>Ongoing constructs</h2><br>435. pSBLbC-CP29-amilCP<br>440. pSBLbC-red<br><br><h2>Todays work</h2><br><b>Plasmid preparation:</b><br>440. pSBLbC-red<br><br><b>Assembly and transformation:</b><br>435. pSBLbC-CP29-amilCP<br></div>' | ||
| + | } | ||
| + | else | ||
| + | { | ||
| + | ds = 'Nothing happend during this day!'; | ||
| + | } | ||
| + | document.getElementById(id).setAttribute('onclick','closeNote('+y+', '+m+', '+d+');'); | ||
| + | document.getElementById(id).innerHTML = ds; | ||
| + | } | ||
| + | |||
| + | function closeNote(y, m, d) | ||
| + | { | ||
| + | var ds = ''; | ||
| + | var id = 'd'+y+m+d; | ||
| + | var ds = '<h1>'+y+'-'+m+'-'+d+'</h1>'; | ||
| + | document.getElementById(id).setAttribute('onclick','diry('+y+', '+m+', '+d+');'); | ||
| + | document.getElementById(id).innerHTML = ds; | ||
| + | } | ||
| + | |||
| + | </script> | ||
| + | </html> | ||
Latest revision as of 23:26, 4 October 2013
