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Template:Team:Uppsala/JS/notebook
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ds = '<div id="dairy-text"><h1>Tuesday 2013-08-13</h1><br><b>Name of participants: </b>Anders Edlund, Mikael Strandgren, Anton Berglund, Viktor Törnblom, Viktor Blomkvist, Christoffer Ahlström, Stephanie Herman, Hampus.E, Karl H, Ken B, Kristoffer L, Victor S, Marcus H, Theodor L. Lovisa P, Alexander, Malin, Pontus, Niclas, Christoffer, Nils, Sabri<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br><br><b>E. coli PK401 dam-:</b><br>111.7 pSBLb4C15-red<br>181. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>114.(2-3) pSB1C3-B0034-TAL-mut<br>115.(1-3) pSB3K3-J23101-B0034-His-TAL<br>204.(1-3) pEL3S17-J23110-B0034-HIS-STS<br><br><h2>Todays work:</h2><br><b>O/N Screening glycerolstock:</b><br>110.pEL3S17-K206001-B0034-IspA<br>219.pSB1C3-Miraculin<br>253.pEL3S17-J23101-B0034-UGTCs2-His<br><br><b>Assembly</b><br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-meffRed à 155. pSB1C3-j23110-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-meffRed à 156. pSB1C3-j23106-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-spisPink à 157. pSB1C3-j23110-RBS-spisPink<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-spisPink à 158. pSB1C3-j23106-RBS-spisPink<br><br><b>Ligation:</b><br>134.1 pSB1C3-CBP (134.1 from digested plasmids [PCR product])<br>134.2 pSB1C3-CBP (134.2 from digested plasmids [PCR product])<br>254.1 pSB1C3-gus (254.1 from digested plasmids [PCR product])<br>254.1 pSB1C3-gus (254.2 from digested plasmids [PCR product])<br>255.1 pSBLb4C15-pemp (257.1 from digest plasmids [PCR product])<br>255.2 pSBLb4C15-pemp (257.2 from digest plasmids [PCR product])<br>256.1 pSBLb4C15-pem (258.1 from digested plasmids [PCR product])<br>256.2 pSBLb4C15-pem (258.2 from digested plasmids [PCR product])<br><br><b>Plasmidprepp:</b><br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>222.1.pSB3K3-CP1-B0034-UGTCs2-His<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His<br><br><b>Sequencing prep:</b><br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>114.(2-3) pSB1C3-B0034-TAL-mut<br>115.(1-3) pSB3K3-J23101-B0034-His-TAL<br>204.(1-3) pEL3S17-J23110-B0034-HIS-STS<br>108.2.pEL3S17-J23106-B0034-IspA<br>108.3.pEL3S17-J23106-B0034-IspA<br>163.1.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>222.1.pSB3K3-CP1-B0034-UGTCs2-His<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His<br><br><b>Re-streaking:</b><br>134. pSB1C3-CBP (CBP1 [134.1] from digested plasmids [PCR-product])<br>134. pSB1C3-CBP (CBP2 [134.2] from digested plasmids [PCR-product])<br>254. pSB1C3-gus (gus1 [254.1] from digested plasmids [PCR-product])<br>254. pSB1C3-gus (gus2 [254.2] from digested plasmids [PCR-product])<br>254. pSB1C3-gus (gus3 [254.3] from digested plasmids [PCR-product])<br>255. pSBLb4C15-pemp (pemp1 [257.1] from digested plasmids [PCR-product])<br>255. pSBLb4C15-pemp (pemp2 [257.2] from digested plasmids [PCR-product])<br>256. pSBLb4C15-pem (pem1 [258.1] from digested plasmids [PCR-product])<br>256. pSBLb4C15-pem (pem2 [258.2] from digested plasmids [PCR-product])<br>256. pSBLb4C15-pem (pem3 [258.3] from digested plasmids [PCR-product])<br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br><b>Re streak 14/8 or 15/8</b><br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br><br><b>Transformation & plating</b><br>134.1 pSB1C3-CBP (134.1 from digested plasmids [PCR product])<br>134.2 pSB1C3-CBP (134.2 from digested plasmids [PCR product])<br>254.1 pSB1C3-gus (254.1 from digested plasmids [PCR product])<br>254.1 pSB1C3-gus (254.2 from digested plasmids [PCR product])<br>255.2 pSBLb4C15-pemp (257.2 from digest plasmids [PCR product])<br>255.1 pSBLb4C15-pemp (257.1 from digest plasmids [PCR product])256.1 pSBLb4C15-pem (258.1 from digested plasmids [PCR product])<br>256.2 pSBLb4C15-pem (258.2 from digested plasmids [PCR product])<br>225. pEL3S17-J23101-B0034-TAL-B0034-4CL-B0034-STS<br>261. pSB3K3-J23110-B0034-HIS-4CL<br>120. pSB3K3-B0034-HIS-4CL<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>177pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>184.pSB1C3-B0034-CrtZ<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br><br><b>Re-streaking</b><br>254. pSB1C3-gus (gus3 [254.3] from digested plasmids [PCR-product]): 1 clone<br>255. pSBLb4C15-pemp (pemp3 [257.3] from digested plasmids [PCR-product]): 4 clones<br><br><b>O/N:</b><br>275. pSBLb4C15-red (dam-)<br>276. pSBLb4C15-CP1-Exp-B0034-Linker-Miraculin-His (dam)<br>277. pSBLb4C15-CP1-Exp-B0034-Linker-Miraculin-His (dam)<br><br><b>Gradient PCR:</b> <br>CBP (colony PCR from Lb.Reuteri)<br>CSP (colony PCR from Lb.Reuteri)<br>Mcherry (from plasmid preparation)<br>256. pSBLb4C15-pem<br><br><h2>Results</h2><br>One of the two gels from SDS page was succesfull, still have problems with the buffers<br><br><h2>Characterization</h2><br><b>Characterization of Resveratrol production through spectroscopy and HPLC</b><br><br><br><b>Western blot</b><br><b>SDS-page on constructs:</b><br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>115. pSB3K3-J23101-B0034-His-TAL<br><br><br><b>Electro transfer over night</b><br><br><h2>Other experiments</h2><br><b>Casting LB-agar plates:</b><br>Chloramphenicol: 85<br></div>' | ds = '<div id="dairy-text"><h1>Tuesday 2013-08-13</h1><br><b>Name of participants: </b>Anders Edlund, Mikael Strandgren, Anton Berglund, Viktor Törnblom, Viktor Blomkvist, Christoffer Ahlström, Stephanie Herman, Hampus.E, Karl H, Ken B, Kristoffer L, Victor S, Marcus H, Theodor L. Lovisa P, Alexander, Malin, Pontus, Niclas, Christoffer, Nils, Sabri<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br><br><b>E. coli PK401 dam-:</b><br>111.7 pSBLb4C15-red<br>181. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>114.(2-3) pSB1C3-B0034-TAL-mut<br>115.(1-3) pSB3K3-J23101-B0034-His-TAL<br>204.(1-3) pEL3S17-J23110-B0034-HIS-STS<br><br><h2>Todays work:</h2><br><b>O/N Screening glycerolstock:</b><br>110.pEL3S17-K206001-B0034-IspA<br>219.pSB1C3-Miraculin<br>253.pEL3S17-J23101-B0034-UGTCs2-His<br><br><b>Assembly</b><br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-meffRed à 155. pSB1C3-j23110-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-meffRed à 156. pSB1C3-j23106-RBS-meffRed<br>pSB1C3-Blue, pSB1C3-J23110, pSB1C3-spisPink à 157. pSB1C3-j23110-RBS-spisPink<br>pSB1C3-Blue, pSB1C3-J23106, pSB1C3-spisPink à 158. pSB1C3-j23106-RBS-spisPink<br><br><b>Ligation:</b><br>134.1 pSB1C3-CBP (134.1 from digested plasmids [PCR product])<br>134.2 pSB1C3-CBP (134.2 from digested plasmids [PCR product])<br>254.1 pSB1C3-gus (254.1 from digested plasmids [PCR product])<br>254.1 pSB1C3-gus (254.2 from digested plasmids [PCR product])<br>255.1 pSBLb4C15-pemp (257.1 from digest plasmids [PCR product])<br>255.2 pSBLb4C15-pemp (257.2 from digest plasmids [PCR product])<br>256.1 pSBLb4C15-pem (258.1 from digested plasmids [PCR product])<br>256.2 pSBLb4C15-pem (258.2 from digested plasmids [PCR product])<br><br><b>Plasmidprepp:</b><br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>222.1.pSB3K3-CP1-B0034-UGTCs2-His<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His<br><br><b>Sequencing prep:</b><br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>114.(2-3) pSB1C3-B0034-TAL-mut<br>115.(1-3) pSB3K3-J23101-B0034-His-TAL<br>204.(1-3) pEL3S17-J23110-B0034-HIS-STS<br>108.2.pEL3S17-J23106-B0034-IspA<br>108.3.pEL3S17-J23106-B0034-IspA<br>163.1.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>142.3.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>222.1.pSB3K3-CP1-B0034-UGTCs2-His<br>222.3.pSB3K3-CP1-B0034-UGTCs2-His<br><br><b>Re-streaking:</b><br>134. pSB1C3-CBP (CBP1 [134.1] from digested plasmids [PCR-product])<br>134. pSB1C3-CBP (CBP2 [134.2] from digested plasmids [PCR-product])<br>254. pSB1C3-gus (gus1 [254.1] from digested plasmids [PCR-product])<br>254. pSB1C3-gus (gus2 [254.2] from digested plasmids [PCR-product])<br>254. pSB1C3-gus (gus3 [254.3] from digested plasmids [PCR-product])<br>255. pSBLb4C15-pemp (pemp1 [257.1] from digested plasmids [PCR-product])<br>255. pSBLb4C15-pemp (pemp2 [257.2] from digested plasmids [PCR-product])<br>256. pSBLb4C15-pem (pem1 [258.1] from digested plasmids [PCR-product])<br>256. pSBLb4C15-pem (pem2 [258.2] from digested plasmids [PCR-product])<br>256. pSBLb4C15-pem (pem3 [258.3] from digested plasmids [PCR-product])<br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br><b>Re streak 14/8 or 15/8</b><br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br><br><b>Transformation & plating</b><br>134.1 pSB1C3-CBP (134.1 from digested plasmids [PCR product])<br>134.2 pSB1C3-CBP (134.2 from digested plasmids [PCR product])<br>254.1 pSB1C3-gus (254.1 from digested plasmids [PCR product])<br>254.1 pSB1C3-gus (254.2 from digested plasmids [PCR product])<br>255.2 pSBLb4C15-pemp (257.2 from digest plasmids [PCR product])<br>255.1 pSBLb4C15-pemp (257.1 from digest plasmids [PCR product])256.1 pSBLb4C15-pem (258.1 from digested plasmids [PCR product])<br>256.2 pSBLb4C15-pem (258.2 from digested plasmids [PCR product])<br>225. pEL3S17-J23101-B0034-TAL-B0034-4CL-B0034-STS<br>261. pSB3K3-J23110-B0034-HIS-4CL<br>120. pSB3K3-B0034-HIS-4CL<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>177pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>184.pSB1C3-B0034-CrtZ<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br><br><b>Re-streaking</b><br>254. pSB1C3-gus (gus3 [254.3] from digested plasmids [PCR-product]): 1 clone<br>255. pSBLb4C15-pemp (pemp3 [257.3] from digested plasmids [PCR-product]): 4 clones<br><br><b>O/N:</b><br>275. pSBLb4C15-red (dam-)<br>276. pSBLb4C15-CP1-Exp-B0034-Linker-Miraculin-His (dam)<br>277. pSBLb4C15-CP1-Exp-B0034-Linker-Miraculin-His (dam)<br><br><b>Gradient PCR:</b> <br>CBP (colony PCR from Lb.Reuteri)<br>CSP (colony PCR from Lb.Reuteri)<br>Mcherry (from plasmid preparation)<br>256. pSBLb4C15-pem<br><br><h2>Results</h2><br>One of the two gels from SDS page was succesfull, still have problems with the buffers<br><br><h2>Characterization</h2><br><b>Characterization of Resveratrol production through spectroscopy and HPLC</b><br><br><br><b>Western blot</b><br><b>SDS-page on constructs:</b><br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>115. pSB3K3-J23101-B0034-His-TAL<br><br><br><b>Electro transfer over night</b><br><br><h2>Other experiments</h2><br><b>Casting LB-agar plates:</b><br>Chloramphenicol: 85<br></div>' | ||
} | } | ||
| - | + | else if(id == 'd2013814') | |
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Wednesday 2013-08-14</h1><br><b>Name of participants: </b>Christoffer A, Stephanie H, Alona N, Mikael S, Viktor B, Viktor T, Alexander, Malin, Pontus, Niclas, Christoffer, Nils, Sabri, Hampus.E, Karl H, Ken B, Kristoffer L, Victor S, Marcus H, Theodor L., Peter C, Lovisa P<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>115. pSB3K3-J23101-B0034-His-TAL<br><br><b>E. coli PK401 dam-:</b><br>111.7 pSBLb4C15-red<br>181. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br><br><h2>Todays work:</h2><br><b>Re-streak:</b><br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>110.pEL3S17-K206001-B0034-IspA<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>pSB1K3-J23110-B0034-aeBlue (should be subcloned to pSB1C3 after)<br>pSB3K3-J23110-B0034-AmilCP (should be subcloned to pSB1C3 after)<br>pSB3K3-J23110-B0034-asCP (should be subcloned to pSB1C3 after)<br>Re streak 15/8 or 16/8<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br><br><b>O/N:</b><br>285. pSB1C3-J23110-B0034-aeBlue<br>286. pSB1C3-J23110-B0034-amilCP<br>287. pSB1C3-J23110-B0034-asCP<br><br><b>Streak from frozen stock:</b><br>111. pSBLb4C15-red<br><br><b>Gradient touchdown colony PCR:</b><br>CSP <br><br><b>Gel Electrophoresis:</b><br>CBP (colony PCR from Lb.Reuteri)<br>CSP (colony PCR from Lb.Reuteri)<br>Mcherry (from plasmid preparation)<br>256. pSBLb4C15-pem<br><br><b>PCR purification: </b><br>M1 <br>M2<br>M3<br>M4<br>CB1<br>CB2<br>256. pSBLb4C15-pem (P1)<br><br><b>Digestion:</b><br>M3 (x+p)<br>CB1 (x+p)<br>P1 (x+p)<br>111.7 pSB4C15-red (e+p)<br>Bleomycin resistance cassette (SacI)<br>pEL PCR product with e. coli ori (ClaI)<br>Lb3 reuteri DSM 20016 pVS2 (ClaI)<br>CP promoters with amilGFP, aeBlue,tsPurple<br><br><b>Ligation:</b><br>M3 with pSB1C3, pSBLB4C15 + CP8, CP1<br>CB1 with pSB1C3<br>P1 with pSB1C3<br>pVS2 origin of replication with pEL PCR-product, cut by ClaI <br>CP promoters with amilGFP, aeBlue,tsPurple<br><br><b>Frozen stock and plasmid preparation:</b><br>275.1 pSBLb4C15-red (dam-)*<br>275.2 pSBLb4C15-red (dam-)<br>276. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His (dam-)<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His (dam-)<br><i>*O/N of clone 1 was made from a very small colony, its colour wasn’t as strong as clone 2’s after growing overnight.</i><br><br><b>Transformation:</b><br>109.pEL3S17-K206000-B0034-IspA x<br>110.pEL3S17-K206001-B0034-IspA x<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His x<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His x<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His x<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His x<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His x<br>184.pSB1C3-B0034-CrtZ x<br>208.pSB1A3-B0034-Thioredoxin-ZCD-His x<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His x<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His x<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His x<br>267.psB3K3-K206000-B0034-IspA x<br>268.psB3K3-K206001-B0034-IspA x<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His x<br>272 pSB1K3-B0034-Thioredoxin-ZCD-His x<br>273 pSB3T5-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His x<br>274 pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His x<br>179 pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His x<br><br><b>Plasmid prepp:</b><br>280. pSB1A3-B0034-aeBlue (should be subcloned to pSB1C3 after, give Erik the subclones version)<br><br><h2>Results:</h2><br><b>Gel Electrophoresis:</b><br>CBP (colony PCR from Lb.Reuteri)<br>CB1: 54.3o C (NC ok!), CB2: 63.3o C (NC ok!) → Correct bands!!<br>CSP (colony PCR from Lb.Reuteri) <br>CS1: 55.7o C (NC ok!), CS2: 65.6o C (NC ok!) → No bands!! (not even primer dimer) <br>Mcherry (from plasmid preparation)<br>M1: 45.0o C (NC ok!), M2: 47.9o C (NC ok!), M3: 50.5o C (NC ok!), M4: 54.3o C (NC ok!) → Correct bands!!<br>256. pSBLb4C15-pem: <br>P1: 47.9o C (NC ok!) → Correct bands!!<br><br><b>PCR purification: </b><br>M1: 65.9 ng/µl → Store in freezer!<br>M2: 200.6 ng/µl → Store in freezer!<br>M3: 136.8 ng/µl → For digest! → Store in freezer!<br>M4: 49.5 ng/µl → Store in freezer!<br>CB1: 90.8 ng/µl → For digest! → Store in freezer!<br>CB2: 56.7 ng/µl → Store in freezer!<br>P1: 56.2 ng/µl → For digest! → Store in freezer!<br><br><b>Plasmid preparation:</b><br>275.1 PK401 dam- pSBLb4C15-red (dam-): 13.5 ng/µl<br>275.2 PK401 dam- pSBLb4C15-red (dam-): 27.9 ng/µl<br>276. PK401 dam- pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His (dam-): 18.3 ng/µl<br>277. PK401 dam- pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His (dam-): 26.6 ng/µl<br><br><b>Western blot:</b><br>Transfer overnight not complete but continued with anyway<br>Very promising results, bands can be seen around 50 kDa but the ladder is pretty weak so we can’t be absolutely sure. Looks like the J23101 promotor does’nt work.<br><br><h2>Characterization experiments</h2><br><b>Characterization of Resveratrol production through spectroscopy and HPLC</b><br><b>Blotting and development of constructs:</b><br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>115. pSB3K3-J23101-B0034-His-TAL<br><br><h2>Other experiments:</h2><br><b>Making MRS-broth (2x600 ml)</b><br><br><b>New buffers for western blot made </b></div>' | ||
| + | } | ||
else if(id == 'd2013919') | else if(id == 'd2013919') | ||
{ | { | ||
Revision as of 17:48, 1 October 2013
