Template:Team:Uppsala/JS/notebook

ds = '<div id="dairy-text"><h1>Thursday 2013-08-08</h1><br><b>Name of participants:</b> Magnus, Pontus, Sabri, Malin, Niclas, Alexander, Nils, Marcus H, Lovisa P, Emil M, Victor S, Hampus.E, Kristoffer L, Malin B, Stephanie H, Mikael S, Anton B<br><br><h2>Ongoing constructs:<h2><br><b>E. coli:</b><br>60. Strain PK401 dam- no plasmid<br>111.7 pSBLb4C15-red<br>139. pSB3K3-J23101.B0032-BFP<br>91. BBa_J61002-J23106<br>92. BBa_J61002-J23110<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>148. pSB1C3-RBS-gfasPurple<br>150. pSB1C3-RBS-spisPink<br>151. pSB1C3-RBS-meffBlue<br>165. pSB1C3-RBS-tsPurple<br>166. pSB1C3-RBS-scOrange<br>167. pSB1C3-RBS-fwYellow<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>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>243. pSB1A2-LacI<br>244. pSB1C3-Promoter-RBS-Bleomysin<br>245. pSB1C3-J23110-B0034-amajCP(amajLime)<br>246. pSB1C3-J23110-b0034-gfasPurple<br>247. pSB1C3-J23110-RBS-tsPurple<br>248. pSB1C3-J23106-RBS-tsPurple<br>249. pSB1C3-J23110-RBS-scOrange<br>250. pSB1C3-J23106-RBS-scOrange<br>251. pSB1C3-J23110-RBS-fwYellow<br>252. pSB1C3-J23106-RBS-fwYellow<br>43.7 pSB1C3-B0034-TAL<br>116. pSB3K3-J23110-B0034-His-TAL<br>117. pSB3K3-CP1-B0034-His-TAL<br>114. pSB1C3-B0034-TAL(MUT)<br>105. pSB3T5-J23101-B0034-TAL<br>107. pSB3T5-CP1-B0034-TAL<br>115. pSB3K5-J23101-B0034-His-TAL<br>119.pSB1C3-B0034-4CL<br>120.pSB1C3-B0034-HIS-4CL<br>137. pSB3K3-CP8-B0034-TAL<br>138. pSB3K3-CP8-B0034-His-TAL<br>139. pSB3K3-J23101-B0034-BFP<br>171. pSB3T5-J23110-B0034-4CL-B0034-STS<br>172. pSB3T5-CP1-B0034-TAL<br>185. pEL3S17-J23110-B0034-TAL-B0034-4CL-B0034-STS<br>186. pEL3S17-CP1-B0034-4CL-B0034-STS<br>187. pEL3S17-CP8-B0034-4CL-B0034-STS<br>188. pEL3S17-J23101-B0034-4CL-B0034-STS<br>189. pSB3K3-J23101-B0034-TAL<br>204. pEL3S17-J23110-HIS-STS<br>205.pSBLB4C15-CP1-B0034-HIS-TAL<br>206.pSB1C3-J23110-B0034-TAL<br>224. pSB1C3-J23101-B0032-TAL<br>225. pEL3S17-J23101-B0032-TAL-B0034-4cl-B0034-STS<br><br><h2>Todays work:</h2><br><b>Transformation:</b><br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>147.pSB1C3-B0034-Thioredoxin (E,P & E,X)<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>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(SKA DET HÄR STÅ HÄR?)<br>no nr. pSB1c3-Thioredoxin<br>no nr. pEL3S17-Thioredoxin<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>Re-streak:</b><br>108.pEL3S17-J23106-B0034-IspA<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.pSB1K3-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>243. pSB1A2-LacI<br>244. pSB1C3-Promoter-RBS-Bleomysin<br>148. pSB1C3-RBS-gfasPurple<br>150. pSB1C3-RBS-spisPink<br>151. pSB1C3-RBS-meffBlue<br>228.pEL3A17-Blue<br>229.pEL3K17-Blue<br>230.pEL3S15-Blue<br>231.pEL3C17-Blue<br><br><bPlasmid prepp</b><br>226. pBAD/AraC-RBS-Luciferas (2012 years kit)<br>227.pBAD/AraC-RBS-Luciferas (2013 years kit)<br>245. pSB1C3-J23110-B0034-amajCP(amajLime)<br>246. pSB1C3-J23110-b0034-gfasPurple<br><br><b>O/N screening:</b><br>109.pEL3S17-K206000-B0034-IspA<br>142.pEL3S17-CP1-B0034-UGTCs2-6xHis<br>143.pEL3S17-CP8-B0034-UGTCs2-6xHis<br>178.pEL3S17-J23110-B0034-Exp-Linker-Miraculin-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>216.pEL3K16-pBAD/AraC-B0034-Jazz-Linker-CrtE-B0034-Blues-Linker-CrtB-B0034-CrtI-Link<br>er-Zif268-B0034-PBSII-Linker-CrtY-B0034-CrtZ-Linker-Gli1<br>219.pSB1C3-Miraculin<br>222.pSB3K3-CP1-B0034-UGTCs2-His<br><br><b>Two step Assembly</b><br>165.1 pSB1C3-RBS-tsPurple, 91. BBa_J61002-J23106 à 247. pSB1C3-J23110-RBS-tsPurple<br>165.1 pSB1C3-RBS-tsPurple, 92. BBa_J61002-J23110 à 248. pSB1C3-J23106-RBS-tsPurple<br>166.1 pSB1C3-RBS-scOrange, 91. BBa_J61002-J23106 à 249. pSB1C3-J23110-RBS-scOrange<br>166.1 pSB1C3-RBS-scOrange, 92. BBa_J61002-J23110 à 250. pSB1C3-J23106-RBS-scOrange<br>167.1 pSB1C3-RBS-fwYellow, 91. BBa_J61002-J23106 à 251. pSB1C3-J23110-RBS-fwYellow<br>167.1 pSB1C3-RBS-fwYellow, 92. BBa_J61002-J23110 à 252. pSB1C3-J23106-RBS-fwYellow<br><br><b>Overnight (screening)</b><br>1. pSB1C3-red<br><br><b>O/N</b><br>60. Strain PK401 dam- no plasmid<br>111.7 pSBLb4C15-red<br><br><b>Streak from frozen stock</b><br>3. pSB3T5-red<br>6. pSB1K3-red<br>42. pSB4S15-red<br>77. pSB4C15-red<br><br><h2>Results:</h2><br><b>One of the SDS-gels failed, resulting in no good gel to stain (falty buffers, continue to search for sources of errors)</b><br><br><b>The succesfull gel was prepared with control buffers.</b><br><br><b>Transformation of promotors with BFP in the shuttle vector, for characterization in Lactobacillus: Some growth, only white colonies, will let them grow overnight again. </b><br><br><h2>Characterization experiments:</h2><br><b>Westernblot </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>119. pSB1C3-B0034-4CL (from 15)<br><br><b>Electro transfer over night</b><br><br><b>Staining of control gel</b><br><br><b>Detection of TAL activity through spectroscopy (day 4)</b></div>'