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Template:Team:Uppsala/JS/notebook
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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>' | ds = '<div id="dairy-text"><h1>Wednesday 2013-08-14</h1><br><b>Name of participants: </b>Christoffer A, Stephanie H, Alona N, Mikael S, Viktor B, Viktor T, Alexander, Malin, Pontus, Niclas, Christoffer, Nils, Sabri, Hampus.E, Karl H, Ken B, Kristoffer L, Victor S, Marcus H, Theodor L., Peter C, Lovisa P<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>115. pSB3K3-J23101-B0034-His-TAL<br><br><b>E. coli PK401 dam-:</b><br>111.7 pSBLb4C15-red<br>181. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br><br><h2>Todays work:</h2><br><b>Re-streak:</b><br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>110.pEL3S17-K206001-B0034-IspA<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>pSB1K3-J23110-B0034-aeBlue (should be subcloned to pSB1C3 after)<br>pSB3K3-J23110-B0034-AmilCP (should be subcloned to pSB1C3 after)<br>pSB3K3-J23110-B0034-asCP (should be subcloned to pSB1C3 after)<br>Re streak 15/8 or 16/8<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br><br><b>O/N:</b><br>285. pSB1C3-J23110-B0034-aeBlue<br>286. pSB1C3-J23110-B0034-amilCP<br>287. pSB1C3-J23110-B0034-asCP<br><br><b>Streak from frozen stock:</b><br>111. pSBLb4C15-red<br><br><b>Gradient touchdown colony PCR:</b><br>CSP <br><br><b>Gel Electrophoresis:</b><br>CBP (colony PCR from Lb.Reuteri)<br>CSP (colony PCR from Lb.Reuteri)<br>Mcherry (from plasmid preparation)<br>256. pSBLb4C15-pem<br><br><b>PCR purification: </b><br>M1 <br>M2<br>M3<br>M4<br>CB1<br>CB2<br>256. pSBLb4C15-pem (P1)<br><br><b>Digestion:</b><br>M3 (x+p)<br>CB1 (x+p)<br>P1 (x+p)<br>111.7 pSB4C15-red (e+p)<br>Bleomycin resistance cassette (SacI)<br>pEL PCR product with e. coli ori (ClaI)<br>Lb3 reuteri DSM 20016 pVS2 (ClaI)<br>CP promoters with amilGFP, aeBlue,tsPurple<br><br><b>Ligation:</b><br>M3 with pSB1C3, pSBLB4C15 + CP8, CP1<br>CB1 with pSB1C3<br>P1 with pSB1C3<br>pVS2 origin of replication with pEL PCR-product, cut by ClaI <br>CP promoters with amilGFP, aeBlue,tsPurple<br><br><b>Frozen stock and plasmid preparation:</b><br>275.1 pSBLb4C15-red (dam-)*<br>275.2 pSBLb4C15-red (dam-)<br>276. pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His (dam-)<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His (dam-)<br><i>*O/N of clone 1 was made from a very small colony, its colour wasn’t as strong as clone 2’s after growing overnight.</i><br><br><b>Transformation:</b><br>109.pEL3S17-K206000-B0034-IspA x<br>110.pEL3S17-K206001-B0034-IspA x<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His x<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His x<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His x<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His x<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His x<br>184.pSB1C3-B0034-CrtZ x<br>208.pSB1A3-B0034-Thioredoxin-ZCD-His x<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His x<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His x<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His x<br>267.psB3K3-K206000-B0034-IspA x<br>268.psB3K3-K206001-B0034-IspA x<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His x<br>272 pSB1K3-B0034-Thioredoxin-ZCD-His x<br>273 pSB3T5-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His x<br>274 pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His x<br>179 pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His x<br><br><b>Plasmid prepp:</b><br>280. pSB1A3-B0034-aeBlue (should be subcloned to pSB1C3 after, give Erik the subclones version)<br><br><h2>Results:</h2><br><b>Gel Electrophoresis:</b><br>CBP (colony PCR from Lb.Reuteri)<br>CB1: 54.3o C (NC ok!), CB2: 63.3o C (NC ok!) → Correct bands!!<br>CSP (colony PCR from Lb.Reuteri) <br>CS1: 55.7o C (NC ok!), CS2: 65.6o C (NC ok!) → No bands!! (not even primer dimer) <br>Mcherry (from plasmid preparation)<br>M1: 45.0o C (NC ok!), M2: 47.9o C (NC ok!), M3: 50.5o C (NC ok!), M4: 54.3o C (NC ok!) → Correct bands!!<br>256. pSBLb4C15-pem: <br>P1: 47.9o C (NC ok!) → Correct bands!!<br><br><b>PCR purification: </b><br>M1: 65.9 ng/µl → Store in freezer!<br>M2: 200.6 ng/µl → Store in freezer!<br>M3: 136.8 ng/µl → For digest! → Store in freezer!<br>M4: 49.5 ng/µl → Store in freezer!<br>CB1: 90.8 ng/µl → For digest! → Store in freezer!<br>CB2: 56.7 ng/µl → Store in freezer!<br>P1: 56.2 ng/µl → For digest! → Store in freezer!<br><br><b>Plasmid preparation:</b><br>275.1 PK401 dam- pSBLb4C15-red (dam-): 13.5 ng/µl<br>275.2 PK401 dam- pSBLb4C15-red (dam-): 27.9 ng/µl<br>276. PK401 dam- pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His (dam-): 18.3 ng/µl<br>277. PK401 dam- pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His (dam-): 26.6 ng/µl<br><br><b>Western blot:</b><br>Transfer overnight not complete but continued with anyway<br>Very promising results, bands can be seen around 50 kDa but the ladder is pretty weak so we can’t be absolutely sure. Looks like the J23101 promotor does’nt work.<br><br><h2>Characterization experiments</h2><br><b>Characterization of Resveratrol production through spectroscopy and HPLC</b><br><b>Blotting and development of constructs:</b><br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>115. pSB3K3-J23101-B0034-His-TAL<br><br><h2>Other experiments:</h2><br><b>Making MRS-broth (2x600 ml)</b><br><br><b>New buffers for western blot made </b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013815') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Thursday 2013-08-15</h1><br><b>Name of participants: </b> Christoffer Ahlström, Stephanie Herman, Alona Nyberg, Mikael Strandgren, Alexander, Malin, Pontus, Niclas, Christoffer, Sabri, Hampus.E, Karl H, Ken B, Kristoffer L, Victor S, Marcus H., Peter C, Lovisa P, Theodor L<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>282.pSB3K3-J23110-B0034-amilCP<br>283.pSB3K3-J23110-B0034-asCCP<br><br><b>E. coli PK401 dam-:</b><br>275.2 pSBLb4C15-red<br>276 pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>292. pSBLb4C15-CP1-mCherry<br>294. pSBLb4C15-CP8-mCherry<br><br><h2>Todays work:</h2><br><b>Digestions:</b><br>Erythromycin resistance cassette from pLUL631 (SacI, Tango buffer)<br>Bleomycin resistance cassette (SalI, FD buffer)<br><br><b>O/N Screening glycerolstock:</b><br>110.pEL3S17-K206001-B0034-IspA<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br><br><b>Re-streak:</b><br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>184.pSB1C3-B0034-CrtZ<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>110.pEL3S17-K206001-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br><br><b>Plasmid prep:</b><br>282.pSB3K3-J23110-B0034-amilCP<br>283.pSB3K3-J23110-B0034-asCCP<br><br><b>Transformation:</b><br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br>292. PK401 dam- pSBLb4C15-CP1-mCherry (dam-)<br>294. PK401 dam- pSBLb4C15-CP8-mCherry (dam-)<br><br><b>Ligations:</b><br>pSBLB4C15 and Bleomycin resistance cassette cut with SacI and SalI<br><br><b>Spreading of strains:</b><br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br><i>E. coli PK401 dam-:</i><br>292. pSBLb4C15-CP1-mCherry<br>294. pSBLb4C15-CP8-mCherry<br><br><b>Screening PCR and gel electrophoresis</b><br><br><h2Results:</h2><br><b>Yesterdays buffers for western blot resulted in smear. </b><br><br><h2>Characterization experiments</h2><br><b>Characterization of Resveratrol production through spectroscopy and HPLC</b><br><br><h2>Other experiments</h2><br><b>Making MRS-agar plates:</b><br>47 Chloramphenicol<br>47 Without antibiotics<br><br><b>Testing yesterdays buffers for western blot.</b><br> <br><b>New separation- and stacking buffers for western blot with a factor 1000 concentration of Tris.</b></div>' | ||
| + | } | ||
| + | else if(id == 'd2013816') | ||
| + | { | ||
| + | ds = '<div id="dairy-text"><h1>Friday 2013-08-16</h1><br><b>Name of participants: </b>Anton Berglund, Stephanie Herman, Mikael Strandgren, Alexander, Pontus, Nils, Christoffer, Sabri, Hampus.E, Kristoffer L, Victor S, Marcus H, Karl H, Theodor L, Peter C<br><br><h2>Ongoing constructs:</h2><br><b>E. coli:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br>134. pSB1C3-CBP<br>139. pSB3K3-J23101-B0034-BFP<br>233. pSBLb4C15-CP1-B0032-BFP<br>234. pSBLb4C15-CP8-B0032-BFP<br>235. pSBLb4C15-CP11-B0032-BFP<br>236. pSBLb4C15-CP29-B0032-BFP<br>237. pSBLb4C15-CP30-B0032-BFP<br>238. pSBLb4C15-CP41-B0032-BFP<br>239. pSBLb4C15-CP44-B0032-BFP<br>254. pSB1C3-gus<br>255. pSBLb4C15-pemp<br>256. pSBLb4C15-pem<br>267 pSB1C3-pemp<br>288. pSB1C3-pem<br>289. pSB1C3-mCherry<br>290. pSB1C3-CBP<br>291. pSBLb4C15-CP1-mCherry<br>293. pSBLb4C15-CP8-mCherry<br>58. pSB1C3- B0034-IspA<br>62.pSB1C3-CP8<br>95. pSB1C3-K206001<br>108.pEL3S17-J23106-B0034-IspA<br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>129.pSB1C3-Idi(mut) <br>140.pSB3T5-J23110-B0034-UGTCs2-6xHis<br>141.pEL3S17-J23101-B0034-UGTCs2-6xHis<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin<br>163.pEL3S17-ZCD-His-B0034-UGTCs2-His<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His<br>179.pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His<br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His<br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>183.pEL3A15-B0034-CrtZ<br>184.pSB1C3-B0034-CrtZ<br>193.pSB1C3-amilGFP<br>194.pSB1C3-B0034-amilCP<br>195.pSB1C3-cjBlue<br>196.pSB1C3-eforRed<br>197.pSB1C3-aeBlue<br>198.pSB1C3-B0034-amilGFP<br>199.pSB1C3-amajCFP<br>200.pSB1C3-B0034-asCP<br>201. E.Coli. Nissle - CrtY-CrtI<br>202.pSB4C5-CrtE-B0034-Blues-CrtB-B0034-CrtI-Linker-Zif268<br>203.pSB1C3-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207.pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>213.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>214.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>215.pSB1K3-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>217.pSB3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>218.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>219.pSB1C3-Miraculin<br>220.pSB3K3-J23110-B0034-UGTCs2-His<br>221.pSB3T5-J23101-B0034-UGTCs2-His<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br>223.pSB3K3-CP8-B0034-UGTCs2-His<br>226. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2012 years kit)<br>227. pSB1C3-pBAD/AraC-RBS-Luciferin (from 2013 years kit)<br>228. pEL3A17-Blue<br>229. pEL3K17-Blue<br>230. pEL3S15-Blue<br>231. pEL3C17-Blue<br>253. pEL3S17-J23101-B0034-UGTCs2-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His<br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His<br>267.psB3K3-K206000-B0034-IspA<br>268.psB3K3-K206001-B0034-IspA<br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His<br>274.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br>282.pSB3K3-J23110-B0034-amilCP<br>283.pSB3K3-J23110-B0034-asCCP<br>225: pEL3S17-J23101-B0034-TAL-B0034-4CL-B0034-STS<br>295: pSB1C3-J23110-TAL-B0034-4Cl-B0034-STS<br><br><b>E. coli PK401 dam-:</b><br>275.2 pSBLb4C15-red<br>276 pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His<br>277. pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His<br>292. pSBLb4C15-CP1-mCherry<br>294. pSBLb4C15-CP8-mCherry<br><br><h2>Todays work:</h2><br><b>O/N Screening & glycerolstock:</b><br>109.pEL3S17-K206000-B0034-IspA<br>110.pEL3S17-K206001-B0034-IspA<br>184.pSB1C3-B0034-CrtZ<br>208. pSB1A3-B0034-Thioredoxin-ZCD-His<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His<br>2 74.pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His<br><br><b>Streak from frozen stock:</b><br>68. pSB3K3-CP1-B0034-BFP<br>69. pSB3K3-CP8-B0034-BFP<br>70. pSB3K3-CP11-B0034-BFP<br>71. pSB3K3-CP29-B0034-BFP<br>72. pSB3K3-CP30-B0034-BFP<br>73. pSB3K3-CP41-B0034-BFP<br>74. pSB3K3-CP44-B0034-BFP<br>111. pSBLb4C15-red<br><br><b>Digestion</b><br>Erythromycin cassette amplicon from pLUL631 (SalI)<br><br><b>Ligation</b><br>pSB1C3-pemp<br>298. pSBLb4E15-red (E-cassette amplicon from pLUL631 with pSBLb4C15 (SacI + SalI))<br>297. pSBLb4B15-red (Bleomycin cassette with pSBLb4C15 (SacI + SalI))<br><br><b>Plasmid preparation:</b><br>204.1 pEL3S17-J23110-B0034-HIS-STS<br>204.3 pEL3S17-J23110-B0034-HIS-STS<br>204.4 pEL3S17-J23110-B0034-HIS-STS<br>254.1 pSB1C3-gus<br>254.2 pSB1C3-gus<br>255.1 pSBLB4C15-pemp<br>255.2 pSBLB4C15-pemp<br>255.3 pSBLB4C15-pemp<br>255.4 pSBLB4C15-pemp<br><br><b>O/N:</b><br>204.1 pEL3S17-J23110-B0034-HIS-STS<br>204.3 pEL3S17-J23110-B0034-HIS-STS<br>204.4 pEL3S17-J23110-B0034-HIS-STS<br>254.1 pSB1C3-gus<br>254.2 pSB1C3-gus<br>255.1 pSBLB4C15-pemp<br>255.2 pSBLB4C15-pemp<br>255.3 pSBLB4C15-pemp<br>255.4 pSBLB4C15-pemp<br>257.1 pSB1C3-pemp<br>257.2 pSB1C3-pemp<br>257.3 pSB1C3-pemp<br>280. pSB1K3-J23110-B0034-aeBlue<br>281. pSB3K3-J23110-B0034-amilCP<br>282. pSB3K3-J23110-B0034-asCP<br><br><b>Transformation:</b><br>297. PK401 dam- pSBLb4B15-red <br>109.pEL3S17-K206000-B0034-IspA <br>110.pEL3S17-K206001-B0034-IspA <br>155pSB1C3-j23110-RBS-meffRed<br>156pSB1C3-j23106-RBS-meffRed<br>157pSB1C3-j23110-RBS-spisPink<br>158pSB1C3-j23106-RBS-spisPink<br>177.pEL3S17-J23101-B0034-Exp-Linker-Miraculin-His <br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-His <br>179 pEL3S17-CP1-B0034-Exp-Linker-Miraculin-His <br>180.pEL3S17-CP8-B0034-Exp-Linker-Miraculin-His <br>181.pSBLb4C15-CP1-B0034-Exp-Linker-Miraculin-His <br>182.pSBLb4C15-CP8-B0034-Exp-Linker-Miraculin-His <br>184.pSB1C3-B0034-CrtZ<br>202pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207pSB4C5-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>208.pSB1A3-B0034-Thioredoxin-ZCD-His <br>247pSB1C3-J23110-RBS-tsPurple<br>248pSB1C3-J23106-RBS-tsPurple<br>249pSB1C3-J23110-RBS-scOrange<br>250pSB1C3-J23106-RBS-scOrange<br>251pSB1C3-J23110-RBS-fwYellow<br>252pSB1C3-J23106-RBS-fwYellow<br>264.pSB3K3-J23101-B0034-Exp-Linker-Miraculin-His <br>265.psB3K3-J23110-B0034-Exp-Linker-Miraculin-His <br>266.psB3K3-CP1-B0034-Exp-Linker-Miraculin-His <br>267.psB3K3-K206000-B0034-IspA <br>268.psB3K3-K206001-B0034-IspA <br>269.psB3K3-CP8-B0034-Exp-Linker-Miraculin-His <br>272 pSB1K3-B0034-Thioredoxin-ZCD-His <br>273 pSB3T5-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His <br>274 pSB3K3-B0034-Thioredoxin-ZCD-His-B0034-UGTCs2-His <br>202. pSB4C5-RBS-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268<br>203. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>207. pSB4C5-RBS-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Linker-Zif268-B0034-PBSII-Linker-CrtY-B0034-HivC-Linker-CrtO-B0034-CrtZ-Linker-Gli1<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple (grow 30 degrees)<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange (grow 30 degrees)<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow (grow 30 degrees)<br>155. pSB1C3-j23110-RBS-meffRed<br>156. pSB1C3-j23106-RBS-meffRed<br>157. pSB1C3-j23110-RBS-spisPink<br>158. pSB1C3-j23106-RBS-spisPink<br>225: pEL3S17-J23101-B0034-TAL-B0034-4CL-B0034-STS<br>295: pSB1C3-J23110-TAL-B0034-4Cl-B0034-STS<br><br><h2>Results:</h2><br><b>Plasmid prep:</b><br>204.1 pEL3S17-J23110-B0034-HIS-STS: 9.7 ng/µl<br>204.3 pEL3S17-J23110-B0034-HIS-STS: 11.9 ng/µl<br>204.4 pEL3S17-J23110-B0034-HIS-STS: 7.6 ng/µl<br>254.1 pSB1C3-gus: 6.7 ng/µl<br>254.2 pSB1C3-gus: 11.5 ng/µl<br>255.1 pSBLB4C15-pemp: 9.9 ng/µl<br>255.2 pSBLB4C15-pemp: 6.3 ng/µl<br>255.3 pSBLB4C15-pemp: 5.0 ng/µl<br>255.4 pSBLB4C15-pemp: 4.6 ng/µl<br><i>Since the concentrations were so low a new set of overnight cultures were prepared.</i><br><br><b>Spectophotometry:</b><br>Inconclusive results.<br><br><h2>Characterization experiments</h2><br><b>Characterization of Resveratrol production through spectroscopy and HPLC</b><br><br><b>Testing of new buffers for western blot.</b><br><br><h2>Other experiments</h2><br><b>Making MRS-broth (2x600 ml)</b><br><br><b>Making LB-agar plates:</b><br>173 Chloramphenicol <i>(chloramphenicol was forgotten in 26 of the plates, but they had already been mixed with 25 others, so also these had to be discarded. 5 plates of doubtful antibiotic content had already been used by the carotenoid group, they were probably ones with ab. E. colis containing pSB3K3 were spread on some of the mixed plates.)</i><br>28 Kanamycin</div>' | ||
} | } | ||
else if(id == 'd2013919') | else if(id == 'd2013919') | ||
Revision as of 18:36, 1 October 2013
