Template:Team:Uppsala/JS/notebook

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ds = '<div id="dairy-text"> <h1>Saturday 2013-07-13</h1>Name of participants: Sabri J, Nils, Kristoffer L, Theodor Löwe, Viktor Blomkvist (VB), Stephanie Herman (SH), Alona Nyberg (AN) <h2>Ongoing constructs:</h2>E.coli (strain D5α): 101. pSB1C3 - DNA Program123456 79. pEL3K16 - CrtE 102. pSB1C3 - CrtE 76. pEL3K16 - CrtI 78. pEL3C18 - CrtY 90 pSB1k3-J23101 91 BBa_J61002-J23106 92 BBa_J61002-J23110 93 BBa_J61002-J23116 44.4 pSB1C3-b0034-His-Tal 44.5 pSB1C3-b0034-His-Tal 111. pSBLb4C15: MluI+NheI 112. pSBLbEc4C15: MluI+ClaI 68. pSB3K3-CP1-B0032-BFP 73. pSB3K3-CP41-B0032-BFP <h2>Todays work</h2>Ligation: 1. pSB1C3 - red → DNA program123456 (PCR prod) 1. pSB1C3 - red → CrtE (PCR prod) 27. pEL3K16 - red → CrtE (PCR prod) PCR and Gelelectrophoresis: CrtO Transformation: 101. pSB1C3 - DNA program123456 102. pSB1C3 - CrtE Overnight, Screening: 76.(1-4) pEL3K16 - CrtI 78.(1-3) pEL3C18 - CrtY PCR purification: 111. pSBLb4C15 112. pSBLbEc4C15 Digestion: 1. pSB1C3 - red → DNA program123456 (PCR prod) 111. pSBLb4C15: MluI+NheI 112. pSBLbEc4C15: MluI+ClaI Lb13. Lc. lactis MG1363-pJP059, Ori: Two experiments, Mlu1+NheI & MLuI+ClaI * Parts of what will become the first shuttle-vector. Plasmid preparation: 68. pSB3K3-CP1-B0032-BFP 73. pSB3K3-CP41-B0032-BFP <h2>Results</h2>Gel: 47. pSB1C3-B0034-STS PCR product with and without DMSO (7%) Good results on GEL, both PCR with and without DMSO looks good. <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> CrtE PCR prod should have been digested with X,P therefore construct 79 and 102 needs to be redone </div>';

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ds = '<div id="dairy-text"> <h1>Friday 2013-07-12</h1>Name of participants: Kristoffer L, Emil M, Karl H, Peter C, Lovisa P, Theodor L, Ken B-A, Christoffer F, Sabri J, Niclas <h2>Ongoing constructs:</h2>E.coli (strain D5α): 15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT 44. pSB1C3-B0034-His-TAL 45. pSB1C3-B0034-4CL 46. pSB1C3-B0034-His-4CL 47. pSB1C3-B0034-STS 48. pSB1C3-B0034-His-STS 82. pSB4S15-CP29-B0032-BFP 84. pSB4S15-CP6 85. pSB4S15-CP25 87. pSB4S15-CP25-B0032-BFP 89. pSB4S15-CP44-B0032-BFP 97. pSB4S15-CP8-B0032-BFP 98. pSB4S15-CP11-B0032-BFP 100. pJP059 Lb. delbrueckii bulgaricus: 18. Lb. delbrueckii bulgaricus Lc. thermophilus: 19. L.thermophilus L. plantarum: 5. 256-rifR-pAMβ1 10. 256-p256 11. 36E-noplasmid E. faecalis: 9. JH2-2 pAMβ1 L. lactis: 13. MG1363-pJP059 15. unknown-pGus 16. MG1363-noplasmid <h2>Todays work</h2>Digestion: 22. B0032-BFP - E,P 11. psB4A15 Ligation: CrtE~I /w RBS + Zincfinger -> 11. pSB4A15-ed CrtE(PCR-prod.) -> 27. pEL3K16-red CrtE -> 1. pSB1C3 (to registry) -all AMP plates from yesterday failed 90-93. (J23 series promotors) Digest of construct 14 and 15 from plasmid prep with Pst1, followed by gel analysis. Transformation: CrtE~I /w RBS + Zincfinger -> 11. pSB4A15-ed CrtE(PCR-prod.) -> 27. pEL3K16-red 82. pSB4S15-CP29-B0032-BFP 84. pSB4S15-CP6 85. pSB4S15-CP25 87. pSB4S15-CP25-B0032-BFP 89. pSB4S15-CP44-B0032-BFP 97. pSB4S15-CP8-B0032-BFP 98. pSB4S15-CP11-B0032-BFP 100. pJP059 J23101 PCR on DNA program 123456 Plasmid prepp on 96.Cs42s clone nr 1 and 2. Glycerol stock on 96.Cs42s clone 1 and 2. Gel run on digest from plasmid preparation of 96.cs42s clone 1 and 2. Re-streak: 76 pEL3K16 - CrtI (4 clones) 78. pEL3C18 - CrtY (3 clones) 74. pSB3K3-CP44-B0032-BFP 81. pSB4S15-CP41-B0032-BFP 83. pSB4S15-CP30-B0032-BFP 86. pSB4S15-CP6-B0032-BFP 88. pSB4S15-CP1-B0032-BFP 73. pSB3K3-CP41-B0032-BFP Gel electrophoresis: 45. pSB1C3-B0034-4CL 46. pSB1C3-B0034-His-4CL 47. pSB1C3-B0034-STS 46. pSB1C3-B0034-His-4CL 48. pSB1C3-B0034-His-STS pSB4C15 PCR-product with intact ori pSB4C15 PCR-product without ori Overnight culture: 68. pSB3K3-CP1-B0032-BFP 73. pSB3K3-CP41-B0032-BFP 74. pSB3K3-CP44-B0032-BFP Lb10.3 plantarum 256 p256 Lb10.4 plantarum 256 p256 Spreading: 87. pSB4S15-CP25-B0032-BFP 89. pSB4S15-CP44-B0032-BFP 97. pSB4S15-CP8-B0032-BFP 82. pSB4S15-CP29-B0032-BFP 98. pSB4S15-CP11-B0032-BFP Lb10.3 plantarum 256 p256 Lb10.4 plantarum 256 p256 <h2>Results</h2>Transformation: Successful: 76 pEL3K16-CrtI 78 pEL3C18-CrtY failed: 75 pEL3A15-CrtB 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. The result from the gel run of the two 96.Cs42s clones indicates an construct with a size of ~4000 bp. Gel on PCRs: Successful: 45. pSB1C3-B0034-4CL 46. pSB1C3-B0034-His-4CL failed 47. pSB1C3-B0034-STS 46. pSB1C3-B0034-His-4CL 48. pSB1C3-B0034-His-STS -PCR-amplification: 47. pSB1C3-B0034-STS, change temperature -2 degrees 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. <h2>Other experiments</h2>Casting LB-agar plates: AB 27 plates containing spectinomycin. Preparation for sequencing: 69.1 pSB3K3-CP8-B0032-BFP 71.3 pSB3K3-CP29-B0032-BFP 72.3 pSB3K3-CP30-B0032-BFP Dilution of primers: 200 µl VF2 (5 µM) 200 µl VR (5 µM) </div>';

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ds = '<div id="dairy-text"> <h1>Thursday 2013-07-11</h1> Name of participants: 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 <h2>Ongoing constructs:</h2> E.coli (strain D5α): 15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT 44. pSB1C3-B0034-His-TAL 45. pSB1C3-B0034-4CL 46. pSB1C3-B0034-His-4CL 47. pSB1C3-B0034-STS 48. pSB1C3-B0034-His-STS 81. pSB4S15-CP41-B0032-BFP 83. pSB4S15-CP30-B0032-BFP 86. pSB4S15-CP6-B0032-BFP 87. pSB4S15-CP25-B0032-BFP 88. pSB4S15-CP1-B0032-BFP 89. pSB4S15-CP44-B0032-BFP 82. pSB4S15-CP29-B0032-BFP 97. pSB4S15-CP8-B0032-BFP 98. pSB4S15-CP11-B0032-BFP 74. pSB3K3-CP44-B0032-BFP 68. pSB3K3-CP1-B0032-BFP 73. pSB3K3-CP41-B0032-BFP 84. pSB4S15-CP6 85. pSB4S15-CP25 96. Cs42s L. plantarum: 10. 256-p256 <h2>Todays work</h2> Transformation: 81. pSB4S15-CP41-B0032-BFP 83. pSB4S15-CP30-B0032-BFP 86. pSB4S15-CP6-B0032-BFP 87. pSB4S15-CP25-B0032-BFP 88. pSB4S15-CP1-B0032-BFP 89. pSB4S15-CP44-B0032-BFP 82. pSB4S15-CP29-B0032-BFP 97. pSB4S15-CP8-B0032-BFP 98. pSB4S15-CP11-B0032-BFP 74. pSB3K3-CP44-B0032-BFP 68. pSB3K3-CP1-B0032-BFP 73. pSB3K3-CP41-B0032-BFP 77. pSB4C15-red* 75. pEL3A15 - CrtB 76 pEL3K16 - CrtI 78. pEL3C18 - CrtY Restreaks: 68. pSB3K3-CP1-B0032-BFP 73. pSB3K3-CP41-B0032-BFP Lb10.3 plantarum 256 p256 * Lb10.4 plantarum 256 p256 Lb10.5 plantarum 256 p256 * Lb10.6 plantarum 256 p256 PCR: 77.(1-4) pSB4C15-red* *Two different amplifications of plasmid adding restriction-sites with different primers. One product with no removals and one without ori. PCR-amplification: 45. pSB1C3-B0034-4CL 46. pSB1C3-B0034-His-4CL 47. pSB1C3-B0034-STS 48. pSB1C3-B0034-His-STS Digest: 14 pET13b(amp)-zifz:4cl-PBSII:STS 15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT -from plasmid prep with Pst1. PCR screening & gel extraction: 44. pSB1C3-B0034-His-TAL 45. pSB1C3-B0034-4CL Ligation: 84. pSB4S15-CP6 85. pSB4S15-CP25 Plasmid preparation: 77. pSB4C15-red PCR screening of cs42s with RedTaq. Overnight culture on transfrormed 96.Cs42s. Prepare of Vf2 and VR primer solution, 20x. 5 tubes each with 200 microliter/tube. Subcloning of the following CrtB, CrtI, CrtY 23. pSB1C3-CrtB → pEL3A15 - red 24. pSB1C3-CrtI → pEL3K16 - red 25. CrtY(PCR prod.) → pEL3C18 - red <h2>Results</h2> Plasmid preparation: 77.1 pSB4C15-red: 47.2 ng/µl 77.2 pSB4C15-red: 50.2 ng/µl 77.3 pSB4C15-red: 44.0 ng/µl 77.4 pSB4C15-red: 54.4 ng/µl Screening: didn’t look good, the pcr product was approximatley 1000 bp on the gel. Should have been around 4000. <h2>Other experiments</h2> Sequence verification: 61.1 pSB1C3-CP1 63.1 pSB1C3-CP11 67.1 pSB1C3-Cp44 Casting LB-agar plates: 26 plates containing spectinomycin. Preparation of LB-agar solution: transformation from kit: 90. BBa_J61002 (amp)-J23101 91. BBa_J61002 (amp)-J23106 92. BBa_J61002 (amp)-J23110 93. BBa_J61002 (amp)-J23116 Restreek from stock: 2012 SD(Strain Database) 13 and 61(promotors). Aliquoting VF2- and VR-primers. </div>';

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ds = '<div id="dairy-text"> <h1>Wednesday 2013-07-10</h1> Name of participants: 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) <h2>Ongoing constructs:</h2> E.coli (strain D5α): 15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT 43. pSB1C3-B0034-TAL 44. pSB1C3-B0034-His-TAL 45. pSB1C3-B0034-4CL 46. pSB1C3-B0034-His-4CL 47. pSB1C3-B0034-STS 48. pSB1C3-B0034-His-STS 96. p?A?-Cs42s 68. pSB3K3-CP1-B0032-BFP 73. pSB3K3-CP41-B0032-BFP 74. pSB3K3-Cp44-B0032-BFP 81. pSB4S15-CP41-B0032-BFP 83. pSB4S15-CP30-B0032-BFP 86. pSB3K3-CP44-B0032-BFP 87. pSB3K3-CP1-B0032-BFP 88. pSB3K3-CP41-B0032-BFP 89. pSB4S15-CP44-B0032-BFP <h2>Todays work</h2> Digestion: 22. pSB1C3-RBS-B0032-BFP Ligation: 44. pSB1C3-B0034-His-TAL 68. pSB3K3-CP1-B0032-BFP 73. pSB3K3-CP41-B0032-BFP 74. pSB3K3-Cp44-B0032-BFP 81. pSB4S15-CP41-B0032-BFP 83. pSB4S15-CP30-B0032-BFP 86. pSB3K3-CP44-B0032-BFP 87. pSB3K3-CP1-B0032-BFP 88. pSB3K3-CP41-B0032-BFP 89. pSB4S15-CP44-B0032-BFP transformation: 44. pSB1C3-B0034-His-TAL 68. pSB3K3-CP1-B0032-BFP 73. pSB3K3-CP41-B0032-BFP 74. pSB3K3-Cp44-B0032-BFP 81. pSB4S15-CP41-B0032-BFP 83. pSB4S15-CP30-B0032-BFP 86. pSB3K3-CP44-B0032-BFP 87. pSB3K3-CP1-B0032-BFP 88. pSB3K3-CP41-B0032-BFP 89. pSB4S15-CP44-B0032-BFP Re-streak 96. p?A?-cs42s -Streak of transformations of ligated 20 and positive controll (AV,NA) PCR of 19(digested EP), 20(digested EP), 21(digested EP) and CrtO(from 21digestEP) (AV,NA) Spreading of strains: Lb4. reuteri 100-23 no plasmid Lb12. reuteri DSM 20016 no plasmid Lb10. plantarum 256 p256 Lb11. plantarum 36E no plasmid Lb10. plantarum 256 noplasmid: Growth Lb11. plantarum 36E noplasmid: Growth → Spread out (1x) on erythromycin plates, to examine whether they are naturally resistant. <h2>Results</h2> Transformation: Lb4. reuteri 100-23 noplasmid: Growth Lb12. reuteri DSM 20016 noplasmid: Growth Re-streak: 96. p?A?-cs42s worked well. O/N culture: 69.1 pSB3K3-CP8-B0032-BFP 69.2 pSB3K3-CP8-B0032-BFP 69.3 pSB3K3-CP8-B0032-BFP 69.4 pSB3K3-CP8-B0032-BFP 70.1 pSB3K3-CP11-B0032-BFP 70.2 pSB3K3-CP11-B0032-BFP 70.3 pSB3K3-CP11-B0032-BFP 70.4 pSB3K3-CP11-B0032-BFP 71.1 pSB3K3-CP29-B0032-BFP 71.2 pSB3K3-CP29-B0032-BFP 71.3 pSB3K3-CP29-B0032-BFP 71.4 pSB3K3-CP29-B0032-BFP 72.1 pSB3K3-CP30-B0032-BFP 72.2 pSB3K3-CP30-B0032-BFP 72.3 pSB3K3-CP30-B0032-BFP 72.4 pSB3K3-CP30-B0032-BFP *Growth Plasmid preparation: 69.1 pSB3K3-CP8-B0032-BFP: 80.5 ng/µl 69.2 pSB3K3-CP8-B0032-BFP: 75.7 ng/µl 69.3 pSB3K3-CP8-B0032-BFP: 60.5 ng/µl 69.4 pSB3K3-CP8-B0032-BFP: 50.8 ng/µl 70.1 pSB3K3-CP11-B0032-BFP: 71.7 ng/µl 70.2 pSB3K3-CP11-B0032-BFP: 56.3 ng/µl 70.3 pSB3K3-CP11-B0032-BFP: 56.0 ng/µl 70.4 pSB3K3-CP11-B0032-BFP: 45.3 ng/µl 71.1 pSB3K3-CP29-B0032-BFP: 54.0 ng/µl 71.2 pSB3K3-CP29-B0032-BFP: 49.1 ng/µl 71.3 pSB3K3-CP29-B0032-BFP: 83.0 ng/µl 71.4 pSB3K3-CP29-B0032-BFP 67.8 ng/µl 72.1 pSB3K3-CP30-B0032-BFP 53.8 ng/µl 72.2 pSB3K3-CP30-B0032-BFP 71.8 ng/µl 72.3 pSB3K3-CP30-B0032-BFP 109.6 ng/µl 72.4 pSB3K3-CP30-B0032-BFP 106.8 ng/µl </div>';

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else if(id == 'd201379')

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ds = '<div id="dairy-text"> <h1>Tuesday 2013-07-09</h1> Name of participants: 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) <h2>Ongoing constructs:</h2> E.coli (strain D5α): 15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT 43. pSB1C3-B0034-TAL 44. pSB1C3-B0034-His-TAL 45. pSB1C3-B0034-4CL 46. pSB1C3-B0034-His-4CL 47. pSB1C3-B0034-STS 48. pSB1C3-B0034-His-STS 96. p?-Cs42s 81. pSB4S15-CP41 83. pSB4S15-CP30 84. pSB4S15-CP6 85. pSB4S15-CP25 86. pSB3K3-CP44 87. pSB3K3-CP1 88. pSB3K3-CP41 89. pSB4S15-CP44 90. pSB4S15-CP1 69. pSB3K3-CP8-B0032-BFP 70. pSB3K3-CP11-B0032-BFP 71. pSB3K3-CP29-B0032-BFP 72. pSB3K3-CP30-B0032-BFP L. reuteri: Lb4. 100-23-no plasmid Lb12. DSM 20016-no plasmid Lb10. 256-noplasmid LB11. 36E-noplasmid <h2>Todays work</h2> Gel analysis: 43. pSB1C3-B0034-TAL 44. pSB1C3-B0034-His-TAL 45. pSB1C3-B0034-4CL 46. pSB1C3-B0034-His-4CL Ligation: 44. pSB1C3-B0034-His-TAL 45. pSB1C3-B0034-4CL OD600 measurement: Lb4. reuteri 100-23 no plasmid Lb12. reuteri DSM 20016 no plasmid Lb10. plantarum 256 no plasmid Lb11. plantarum 36E no plasmid Competent cells preparation: Lb4. reuteri 100-23 no plasmid Lb12. reuteri DSM 20016 no plasmid Lb10. plantarum 256 no plasmid Lb11. plantarum 36E no plasmid Transfromation: Lb4. reuteri 100-23 no plasmid Lb12. reuteri DSM 20016 no plasmid Lb10. plantarum 256 no plasmid Lb11. plantarum 36E no plasmid Re-streaks: 77. pSB4C15-red Transformation: 81. pSB4S15-CP41 83. pSB4S15-CP30 84. pSB4S15-CP6 85. pSB4S15-CP25 86. pSB3K3-CP44 87. pSB3K3-CP1 88. pSB3K3-CP41 89. pSB4S15-CP44 90. pSB4S15-CP1 44. pSB1C3-B0034-His-TAL 45. pSB1C3-B0034-4CL O/N culture: 69.1 pSB3K3-CP8-B0032-BFP 69.2 pSB3K3-CP8-B0032-BFP 69.3 pSB3K3-CP8-B0032-BFP 69.4 pSB3K3-CP8-B0032-BFP 70.1 pSB3K3-CP11-B0032-BFP 70.2 pSB3K3-CP11-B0032-BFP 70.3 pSB3K3-CP11-B0032-BFP 70.4 pSB3K3-CP11-B0032-BFP 71.1 pSB3K3-CP29-B0032-BFP 71.2 pSB3K3-CP29-B0032-BFP 71.3 pSB3K3-CP29-B0032-BFP 71.4 pSB3K3-CP29-B0032-BFP 72.1 pSB3K3-CP30-B0032-BFP 72.2 pSB3K3-CP30-B0032-BFP 72.3 pSB3K3-CP30-B0032-BFP 72.4 pSB3K3-CP30-B0032-BFP 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. Ligation of 20 with all diffrent combinations of ligase buffer and ligase. (AV, CF, NA) 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) <h2>Results</h2> O/N: Lb4. reuteri 100-23 no plasmid (clone 1): NC ok! → follow protocol for transformation Lb12. reuteri DSM 20016 no plasmid (clone 1): NC ok! → follow protocol for transformation, watch in microscope for contamination Lb10. plantarum 256 no plasmid (clone 1): NC ok! → follow protocol for transformation Lb11. plantarum 36E no plasmid (clone 1): NC ok! → follow protocol for transformation Transformation: 77. pSB4C15-red: Growth on Cm, some red colonies, some blue, many white (?!) OD600 measurement: Lb4. reuteri 100-23 no plasmid: 0.195 (Measure) & 0.668 (Cultivate) → proceed Lb12. reuteri DSM 20016 no plasmid: 0.179 (Measure) & 0.680 (Cultivate) --”-- Lb10. plantarum 256 no plasmid: 0.115 (Measure) & 0.724 (Cultivate) --”-- Lb11. plantarum 36E no plasmid: 0.148 (Measure) & 0.624 (Cultivate) --”-- Microscopy: Lb12. reuteri DSM 20016 no plasmid: No contamination was sighted! → proceed! transformation: Successful: 45. pSB1C3-B0034-4CL 96. p?-Cs42s failed: 44. pSB1C3-B0034-His-TAL <h2>Other experiments</h2>Cold buffer preparation (1 l) 50% glycerol stock preparation (100 ml) Microscopy: Lb12. reuteri DSM 20016 no plasmid </div>';

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else if(id == 'd201378')

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{

+

ds = '<div id="dairy-text"><h1>Monday 2013-07-08</h1> Name of participants: Alexander, Nils, Christofffferz , Magnus, Niclas, Thorsteinn, Kristoffer L, Emil M, Marcus H, Karl H, Peter C, Ken B.-A., Lovisa P <h2>Ongoing constructs</h2> E.coli (strain D5α): 15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT 43. pSB1C3-B0034-TAL 44. pSB1C3-B0034-His-TAL 45. pSB1C3-B0034-4CL 46. pSB1C3-B0034-His-4CL 47. pSB1C3-B0034-STS 48. pSB1C3-B0034-His-STS 57. pSB1C3-B0034-Idi 58. pSB1C3-B0044-IspA 81. pSB4S15-CP41 83. pSB4S15-CP30 84. pSB4S15-CP6 85. pSB4S15-CP25 86. pSB3K3-CP44 87. pSB3K3-CP1 88. pSB3K3-CP41 89. pSB4S15-CP44 90. pSB4S15-CP1 L. reuteri: Lb4. 100-23-no plasmid Lb12. DSM 20016-no plasmid L. plantarum: Lb10. 256-noplasmid LB11. 36E-noplasmid <h2>Todays work</h2> PCR amplification and gel analysis: 43. pSB1C3-B0034-TAL 44. pSB1C3-B0034-His-TAL 45. pSB1C3-B0034-4CL 46. pSB1C3-B0034-His-4CL Plasmid prep.: 47. pSB1C3-B0034-STS Clone: 47.9 and 47.20 15. pSB4K5-RBS-TAL-Linker-Zif268-RBS-4cl-Linker-PBSII-STS-OMT Sequencing prep.: 47. pSB1C3-B0034-STS Clone: 47.9 and 47.20 48. pSB1C3-B0034-His-STS Clone: 48.1 Gel extraction: 43. pSB1C3-B0034-TAL 44. pSB1C3-B0034-His-TAL 45. pSB1C3-B0034-4CL 46. pSB1C3-B0034-His-4CL Restreak: 69. pSB3K3-CP8-B0032-BFP 70. pSB3K3-CP11-B0032-BFP 71. pSB3K3-CP29-B0032-BFP 72. pSB3K3-CP30-B0032-BFP Digest PCR products from succefull gel extraction. Ligation: 81. pSB4S15-CP41 83. pSB4S15-CP30 84. pSB4S15-CP6 85. pSB4S15-CP25 86. pSB3K3-CP44 87. pSB3K3-CP1 88. pSB3K3-CP41 89. pSB4S15-CP44 90. pSB4S15-CP1 Frozen stock: Lb4. reuteri 100-23 no plasmid (clone 1 & 2) Lb12. reuteri DSM 20016 no plasmid (clone 1 & 2) Lb10. plantarum 256 noplasmid (clone 1 & 2) Lb11. plantarum 36E no plasmid (clone 1 & 2) O/N: Lb4. reuteri 100-23 no plasmid (clone 1) Lb12. reuteri DSM 20016 no plasmid (clone 1) Lb10. plantarum 256 no plasmid (clone 1) Lb11. plantarum 36E no plasmid (clone 1) Screening: 43. pSB1C3-B0034-TAL O/N culture: 43.7. pSB1C3-B0034-TAL Subcloning: 23->4, 24->4, 25->4(kontroll), 23->38, 24->27, 25->51 (CF, AV) Transformation of 23->4, 24->4, 25->4(kontroll), 23->38, 24->27, 25->51 (CF, AV) Primer design and order of lambda red (NA) Sequencing preparation of 57.3 and 57.5 (pSB1C3-B0034-Idi), 58.1 and 58.4 (pSB1C3-B0034-IspA) <h2>Results</h2> Microscopy: * *No contamination was seen, save plates for later. → Proceed with O/N of two clones from the no-plasmid strains. Primers for lambda red and Cs42s were ordered today :D. 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) PCR amplification: 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. gel extraction: Successfull 44. pSB1C3-B0034-His-TAL 45. pSB1C3-B0034-4CL failed 43. pSB1C3-B0034-STS 46. pSB1C3-B0034-His-4CL Screening: one succefull clone - 43.7 pSB1C3-B0034-STS <h2>Other experiments</h2> Microscopy: CA* *Of plates from 04/07 and O/N from 07/07 to see if contaminated. </div>';

}

else if(id == 'd201372')

@@ Line 280: / Line 280: @@
    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')
+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 == 'd201372')

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