"
Template:Team:Uppsala/JS/notebook
From 2013.igem.org
(Difference between revisions)
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ds ='<p>på dig</p>'; | ds ='<p>på dig</p>'; | ||
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| - | + | 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>'+ | 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:</h3>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>'+ | '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>'+ | '<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>'+ | ||
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else if(id == 'd2013720') | 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>< | + | 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:</h3><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>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</h3><br><br><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</h3><br><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') | 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>< | + | 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:</h3><br><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</h3><br><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:</h3><br><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') | 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>< | + | 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:</h3><br><br><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</h3><br><br><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</h3><br><br><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') | 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>< | + | 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:</h3><br><br><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</h3><br><br><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</h3><br><br>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') | 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>< | + | 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:</h3><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</h3><br><br><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</h3><br><br><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</h3><br>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') | 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>< | + | 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:</h3><br><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>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</h3><br><br><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</h3><br><br><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 == 'd201372') | else if(id == 'd201372') | ||
| Line 279: | Line 279: | ||
else if(id == 'd201381') | 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>< | + | 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:</h3><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>'+ | '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>'+ | '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>'+ | '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</h3><br><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:</h3><br>185.9-185.10<br>'+ |
| - | '< | + | '<h2>Transformation of constructs:</h3><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</h3><br>Construct 185, clone 9 and 10, bonds around 3000/4000 bp.<br><br>'+ |
| - | '< | + | '<h2>Follow up</h3><br>- <br><br>'+ |
| - | '< | + | '<h2>Other experiments</h3><br>-<br></p></div>'; |
} | } | ||
else if(id == 'd2013919') | else if(id == 'd2013919') | ||
Revision as of 16:16, 17 September 2013
