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ds = '<div id="dairy-text"> <h1>Monday 2013-07-15</h1> Name of participants: Karl H, Lovisa P, Theodor L, Ken B-A, Victor S, Hampus.E, Christoffer, Sabri, Alexander, Nils, Thorsteinn, Magnus, Malin, Pontus, 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 105. pSB3T5-J23101-B0034-TAL 104. pSB1C3-B0034-4CL-B0034-STS 106. pSB3T5-J23110-B0034-TAL 107. pSB3T5-CP1-B0034-TAL 79. pEL3K16 - CrtE 102. pSB1C3 - CrtE 75. pEL3A15 - CrtB 101. pSB1C3 - DNA program123456 76. pEL3K16-CrtI 78. pEL3C18-CrtY 99. pSB1C3 - CrtO 57.Psb1C3-B0034-Idi 91. BBa_J61002 (amp)-J23106 94. pSB1C3-K206000 95. pSB1C3-K206001 11. pSB4A15-blue(?) 58. pSB1C3-IspA 108. pSB4A15-J23106-B0034-IspA 109. pSB4A15-K206000-B0034-IspA 110. pSB4A15-K206001-B0034-IspA 7.pSB3k3-red 22. B0032-BFP 42. pSB4S15-red 60. PK401 dam 61. pSB1C3-CP1 62. pSB1C3-CP8 63. pSB1C3-CP11 64. pSB1C3-CP29 65. pSB1C3-CP30 66. pSB1C3-CP41 67. pSB1C3-Cp44 68. pSB3K3-CP1-B0032-BFP 69. pSB3K3-CP8-B0032-BFP 70. pSB3K3-CP11-B0032-BFP 71. pSB3K3-CP29-B0032-BFP 72. pSB3K3-CP30-B0032-BFP 73. pSB3K3-CP41-B0032-BFP 74. pSB3K3-CP44-B0032-BFP 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 97. pSB4S15-CP8-B0032-BFP 82. pSB4S15-CP29-B0032-BFP 98. pSB4S15-CP11-B0032-BFP 100. pJP059 111. pSBLb4C15 112. pSBLbEc4C15 L. reuteri: 1. CF48-pLR581 2. 1063-pLUL631 3. DSM 20016-pVs2 4. 100-23-noplasmid 6. DSM 20016-pLUL63TsA8 7. DSM 20016-pGFP 8. 100-23-pGT232 14. DSM 20016-pLUL631(B?) 22. Betacarotene-various plasmids L. plantarum: 5. 256-rifR-pAMβ1 10. 256-p256 <h2>Todays work</h2>PCR Purification: 111. pSBLb4C15, digest 112. pSBLbEc4C15, digest Ligation: 111. pSBLb4C15 (MluI+NheI) + Ori from pJP059 (MluI+NheI) 112. pSBLbEc4C15: (MluI+ClaI) + Ori from pJP059 (MluI+ClaI) Transformation: 111. pSBLb4C15 112. pSBLbEc4C15 75. pEL3A15 - CrtB 99. pSB1c3 - CrtO 108. pSB4A15-J23106-B0034-IspA 109. pSB4A15-K206000-B0034-IspA 110. pSB4A15-K206001-B0034-IspA 45. pSB1C3-B0034-4CL 46. pSB1C3-B0034-His-4CL 47. pSB1C3-B0034-STS 104. pSB1C3-B0034-4CL-B0034-STS 105. pSB3T5-J23101-B0034-TAL 106. pSB3T5-J23110-B0034-TAL 107. pSB3T5-CP1-B0034-TAL Overnight: 74. pSB3K3-CP44-B0032-BFP 101. pSB1C3 - DNA program123456 Fluorescence(BFP) experiment with UV: 68. pSB3K3-CP1-B0032-BFP 69. pSB3K3-CP8-B0032-BFP 70. pSB3K3-CP11-B0032-BFP 71. pSB3K3-CP29-B0032-BFP 72. pSB3K3-CP30-B0032-BFP 73. pSB3K3-CP41-B0032-BFP 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 89. pSB4S15-CP44-B0032-BFP Plasmid preparation: Lb10.1 plantarum 256 p256 Lb10.2 plantarum 256 p256 Lb10.3 plantarum 256 p256 Lb22.1 reuteri betacarotene (strain name?) various plasmids 44.5 pSB1C3-b0034-His-Tal 74. pSB3K3-CP44-B0032-BFP PCR: 48. pSB1C3-B0034-His-STS Gel electhrophoresis 48. pSB1C3-B0034-His-STS Assembly: 105. pSB3T5-J23101-B0034-TAL 106. pSB3T5-J23110-B0034-TAL 107. pSB3T5-CP1-B0034-TAL Re-streak: 79. pEL3K16 - CrtE 102. pSB1C3 - CrtE 75. pEL3A15 - CrtB PCR cloning of: 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 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 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 CrtZ (PCR is done to clone and biobrick CrtZ from the following template, 20. pBAD/AraC-CrtE~CrtZ /w RBS, Zinc Finger Mutagenesis 57. pSB1C3-B0034-Idi Sequence preperation 78.1 pEL3C18-CrtY (VF and VR) 78.3 pEL3C18-CrtY (VF and VR) Screening: 111. pSBLb4C15 112. pSBLbEc4C15 81. pSB4S15-CP41-B0032-BFP 83. pSB4S15-CP30-B0032-BFP 86. pSB4S15-CP6-B0032-BFP 88. pSB4S15-CP1-B0032-BFP 89. pSB4S15-CP44-B0032-BFP 101. pSB1C3 - DNA program123456 Touchdown PCR with Phusion polymerase (signal peptides): CA Collagen binding protein → cloned from strain 14.4 Cell surface protein → cloned from strain 3.1 usp45 → cloned from strain 13.1 <h2>Results</h2>PCR, Succes on: 48. pSB1C3-B0034-His-STS Transformation 45. pSB1C3-B0034-4CL, failed 46. pSB1C3-B0034-His-4CL, failed 104. pSB1C3-B0034-4CL-B0034-STS, failed 106. pSB3T5-J23110-B0034-TAL, failed 108. pSB4A15-J23106-B0034-IspA, failed 109. pSB4A15-K206000-B0034-IspA, failed 110. pSB4A15-K206001-B0034-IspA, failed 111. pSBLb4C15 112. pSBLbEc4C15 -(45,46,104,106) Probably due to weak competent cells -(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. Nanodrop measurement of 57.Psb1C3-B0034-Idi was 80 ng/microliter. Gel: 21 pSB4C5-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268, fail Fluorescence(BFP) experiment with UV: 68. pSB3K3-CP1-B0032-BFP : Glowing (68.2 68.4) 69. pSB3K3-CP8-B0032-BFP : Glowing (69.3 69.4) 70. pSB3K3-CP11-B0032-BFP : Glowing (70.3) 71. pSB3K3-CP29-B0032-BFP : Glowing (71.1 71.2) 72. pSB3K3-CP30-B0032-BFP : Glowing (72.1 72.2) 73. pSB3K3-CP41-B0032-BFP : Glowing (73.4) 74. pSB3K3-CP44-B0032-BFP : Glowing (74.1.1 74.1.2) 81. pSB4S15-CP41-B0032-BFP : Not glowing 83. pSB4S15-CP30-B0032-BFP : Not glowing 86. pSB4S15-CP6-B0032-BFP : Not glowing 88. pSB4S15-CP1-B0032-BFP : Not glowing 89. pSB4S15-CP44-B0032-BFP : Not glowing Plasmid preparation: Lb10.1 plantarum 256 p256: 124.7 ng/µl Lb10.2 plantarum 256 p256: 23.5 ng/µl Lb10.3 plantarum 256 p256: 270.9 ng/µl Lb22.1 reuteri betacarotene (strain name?) various plasmids: 51.4 ng/µl </div>';

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else if(id == 'd2013714')

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ds = '<div id="dairy-text"><h1>Sunday 2013-07-14</h1>Name of participants: Anders Edlund (AE), Viktor Blomkvist (VB), Lovisa P, Christoffer F, Sabri J <h2>Ongoing constructs:</h2>E.coli (strain D5α): 7.pSB3k3-red 22. B0032-BFP 42. pSB4S15-red 60. PK401 dam 61. pSB1C3-CP1 62. pSB1C3-CP8 63. pSB1C3-CP11 64. pSB1C3-CP29 65. pSB1C3-CP30 66. pSB1C3-CP41 67. pSB1C3-Cp44 68. pSB3K3-CP1-B0032-BFP 69. pSB3K3-CP8-B0032-BFP 70. pSB3K3-CP11-B0032-BFP 71. pSB3K3-CP29-B0032-BFP 72. pSB3K3-CP30-B0032-BFP 73. pSB3K3-CP41-B0032-BFP 74. pSB3K3-CP44-B0032-BFP 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 97. pSB4S15-CP8-B0032-BFP 82. pSB4S15-CP29-B0032-BFP 98. pSB4S15-CP11-B0032-BFP 100. pJP059 111. pSBLb4C15 112. pSBLbEc4C15 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 91. BBa_J61002-J23106 92. BBa_J61002-J23110 93. BBa_J61002-J23116 103. pSB1k3-J23101 79. pEL3K16 - CrtE 102. pSB1C3 - CrtE 75. pEL3A15 - CrtB 101. pSB1C3 - DNA program123456 76. pEL3K16-CrtI 78. pEL3C18-CrtY L. reuteri: 1. CF48-pLR581 2. 1063-pLUL631 3. DSM 20016-pVs2 4. 100-23-noplasmid 6. DSM 20016-pLUL63TsA8 7. DSM 20016-pGFP 8. 100-23-pGT232 14. DSM 20016-pLUL631(B?) 22. Betacarotene-various plasmids L. plantarum: 5. 256-rifR-pAMβ1 10. 256-p256 <h2>Todays work</h2> Ligation: 111. pSBLb4C15 (MluI+NheI) + Ori from pJP059 (MluI+NheI) 112. pSBLbEc4C15: (MluI+ClaI) + Ori from pJP059 (MluI+ClaI) Transformation: 111. pSBLb4C15 112. pSBLbEc4C15 79. pEL3K16 - CrtE 102. pSB1C3 - CrtE 75. pEL3A15 - CrtB Plasmid prep. and glycerol stock: 103. pSB1k3-J23101 91. BBa_J61002-J23106 92. BBa_J61002-J23110 93. BBa_J61002-J23116 44.4 pSB1C3-b0034-His-Tal 76.(1-4)-pEL3K16-CrtI 78.(1-3)-pEL3C18-CrtY Overnight: 44.5 pSB1C3-b0034-His-Tal BBa_J61002-J23101 (amp) Restreak: 101. pSB1C3 - DNA program123456 (Picked 4 clones) Subcloning: CrtE (PCR prod) should be digested with X,P (Can be found in digest box) 27. pEL3K16 - red should be digested with X,P → CrtE (PCR prod) 1. pSB1C3 - red should be digested with X,P → CrtE (PCR prod) 38. pEL3A15 - red → 23. pSB1C3-CrtB PCR cloning: 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 CrtZ (PCR is done to clone and biobrick CrtZ from the following template, 20. pBAD/AraC-CrtE~CrtZ /w RBS, Zinc Finger 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 <h2>Results</h2> Problems with the yesterday ligation, the experiment was redone Transformation of 101. DNA program123456 was a sucess although 102. pSB1C3-CrtE failed The PCR failed for CrtE~I, unspecific binding The PCR failed for CrtZ, unspecific binding. </div>';

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else if(id == 'd2013713')

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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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else if(id == 'd2013712')

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{

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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 <h2>Ligation:</h2> 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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else if(id == 'd201372')

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   {
 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><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>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><br>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><br><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> <br><br><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><br><br><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><br><br><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><br><br><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><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>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><br>Digestion:<br>22. B0032-BFP - E,P<br>11. psB4A15<br><br><h2>Ligation:</h2> <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><br><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><br>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 == 'd201372')

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