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==Sep 22== ===Colony PCR=== <div class="experiment"> <span class="author">Tatsui</span> {| class="wikitable" !Sample||base pair |- |9/20 Plac-PT181 attenuator(1~4)||664 |- |9/20 Plac-PT181 antisense(1~6)||369 |- |9/20 Pcon-PT181 attenuator-aptamer-DT(1~6)||859 |- |9/20 Pcon-PT181 antisense-spinach-DT(1~8)||723 |} </div> [[File:Igku_0903_E1.jpg]]<br> [[File:Igku_0922_E2.jpg]]<br> ===Electrophoresis=== <div class="experiment"> <span class="author">Nakamoto</span> {| class="wikitable" !Lane||Sample |- |1||Pcon-apt12_1R-DT(E-1A) |- |2||Pcon-spinach-DT(E-1A) |- |3||Pcon-pT181 attenuator(E-1A) |- |4||DT(1-3) |- |5||tetR aptamer 12_1R-DT(1-3) |- |6||100bp ladder |- |7||Pcon-pT181 attenuator(1-SA) |- |8||spinach-DT(2-1) |- |9||Pcon-pT181 attenuator(E-0) |- |10||Pcon-pT181 attenuator(2-SA) |- |11||spinach-DT(3-2) |- |12||Ptet-PT181 anisense |} [[File:Igku_0922_E3.jpg]]<br> [[File:Igku_0922_E4.jpg]]<br> ===Miniprep=== <div class="experiment"> <span class="author">Honda</span> {|class="wikitable" !DNA||concentration[&micro;g/mL]||260/280||260/230 |- |9/21 spinach-DT||197.9||1.67||1.54 |- |9/21 Plux||125.9||1.71||1.27 |- |9/21 Plux-RBS-GFP-DT-(1)||168.0||1.62||1.73 |- |9/21 Pcon-PT181 attenuator-(1)||368.3||1.57||1.62 |- |9/21 RBS-GFP-DT||532.8||1.75||1.78 |- |9/21 Pcon-RBS-GFP-DT||845.2||2.02||1.70 |} </div> ===Colony PCR=== <div class="experiment"> <span class="author">Honda</span> {| class="wikitable" !Sample||base pair |- |9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1~2)||283 |} {| class="wikitable" !PreDenature||Denature||Annealing||Extension||cycle |- |94&deg;C||94&deg;C||55&deg;C||68&deg;C||-- |- |5min||30s||30s||2min10s||30cycles |} [[File:igku_0922_E5.jpg]]<br> [[File:igku_0922_E6.jpg]]<br> ===Gel Extraction=== <div class="experiment"> <span class="author">No name</span> {| class="wikitable" !Lane||DNA||Enzyme |- |1||100bp ladder||-- |- |2~4||Pcon-aptamer12-1R-DT(E-1A)||-- |- |6~8||Pcon-spinach-DT(E-1A)||-- |- |10~12||Pcon-attenuator PT181(E-1A)||-- |} {| class="wikitable" !Lane||DNA||Enzyme |- |1||100bp ladder||-- |- |2~4||DT(1-3)||-- |- |6~8||aptamer12-1RDT(1-3)||-- |- |10~12||Pcon-PT181 attenuator(1-5A)||-- |} {| class="wikitable" !Name||concentration[&micro;g/mL]||260/280||260/230 |- |Pcon-aptamer12-1R-DT(E-1A)||31.2||1.87||0.30 |- |Pcon-spinach-DT(E-1A)||25.6||1.88||0.99 |- |Pcon-attenuator PT181(E-1A)||25.3||1.99||0.86 |- |DT(1-3)||22.1||1.82||0.20 |- |aptamer12-1RDT(1-3)||33.5||1.99||0.05 |- |Pcon-PT181 attenuator(1-5A)||36.9||1.12||0.96 |- |} </div> ===digestion=== <div class="experiment"> <span class="author">No name</span> {| class="wikitable" ! ||9/16 Plac||XbaI||SpeI||PstI||BufferB||BSA||MilliQ||total |- |2cuts||12.5&micro;L||0&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||9.5&micro;L||30&micro;L |- |NC||0.8&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.4&micro;L||10&micro;L |- |} {| class="wikitable" ! ||9/22 Plux||XbaI||SpeI||PstI||BufferB||BSA||MilliQ||total |- |2cuts||16&micro;L||0&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14&micro;L||30&micro;L |- |NC||0.8&micro;L||0&micro;L||||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.2&micro;L||10&micro;L |} {| class="wikitable" ! ||9/20 RBS-tetR-2R-DT ||XbaI||SpeI||PstI||BufferD||BSA||MilliQ||total |- |2cuts||8.5&micro;L||1&micro;L||0&micro;L||1&micro;L||3&micro;L||3&micro;L||13.5&micro;L||30&micro;L |- |NC||0.4&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.6&micro;L||10&micro;L |- |} ===Gel Extraction=== <div class="experiment"> <span class="author">No name</span> {| class="wikitable" !Lane||DNA||Enzyme |- |1||100bp ladder||-- |- |2~4||Pcon-attenuator PT181(E-0)||-- |} {| class="wikitable" !Lane||DNA||Enzyme |- |1||100bp ladder||-- |- |2~4||Pcon-attenuator PT181(E-0)||-- |} {| class="wikitable" !Lane||DNA||Enzyme |- |2||100bp ladder||-- |- |4~6||Pcon-attenuator PT181(2-SA)||-- |} {| class="wikitable" !Lane||DNA||Enzyme |- |1||100bp ladder||-- |- |3~5||spinach-DT(3-2)||-- |- |8~10||Ptet-antisense PT181(1-3)||-- |} </div> {| class="wikitable" !Name||concentration[&micro;g/mL]||260/280||260/230 |- |Pcon-attenuator PT181(E-0)||2.4||1.80||0.17 |- |Pcon-attenuator PT181(2-SA)||10.8||1.85||0.54 |- |spinach-DT(3-2)||36.7||1.80||0.75 |- |Ptet-antisense PT181(1-3)||16.8||1.74||0.78 |} ===Colony PCR=== <div class="experiment"> <span class="author">No name</span> {| class="wikitable" !Sample||base pair |- |9/20 Pcon-PT181 attenuator-aptamer12-1R-DT-(7~14)||859 |} ===Colony PCR=== <div class="experiment"> <span class="author">Tatsui</span> {| class="wikitable" !Sample||base pair |- |PconPT181antisense-spinach-DT(9~15)||723 |} ===Ligation=== <div class="experiment"> <span class="author">No name</span> {| class="wikitable" !state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2 |- |experiment||9/13 Pcon-PT181 attenuator (SpeI & PstI)||1.7 &micro;L||9/18 aptamer-1R-DT (XbaI & PstI)||5.4&micro;L|| 3.5&micro;L |- |experiment||9/12 Plac(SpeI & PstI)||2.4 &micro;L||9/18 aptamer-1R-DT (XbaI & PstI)||6.1&micro;L||4.5 &micro;L |- |experiment||9/13 Ptet(SpeI & PstI)||1.7 &micro;L||9/10 Plux-RBS-GFP-DT||7.0 &micro;L||4.4 &micro;L |- |experiment||9/21 RBS-GFP-DT (EcoRI & XbaI)||0.6 &micro;L||9/18 aptamer 12-1R-DT (XbaI & PstI)||5.6&micro;L||3.1 &micro;L |- |experiment||9/13 PSB1C3(XbaI & PstI)||1.9 &micro;L||9/16 PT181 attenuator(XbaI & PstI)||4.6&micro;L||3.3 &micro;L |- |} </div> ===Liquid Culture=== <div class="experiment"> <span class="author">No name</span> {| class="wikitable" !Sample||medium |- |9/20 Plac-antisense-1||-- |- |9/20 Plac-attenuator-1||-- |- |9/20 Plac-antisense-1||-- |- |9/20 Plac-attenuator-1||-- |} </div> ===digestion=== <div class="experiment"> <span class="author">No name</span> {| class="wikitable" ! ||9/8 Pcon-PT181 antisense||EcoRI||SpeI|| BufferM||MilliQ||total |- |2cuts||8.3&micro;L||1&micro;L||1&micro;L||3&micro;L||16.7&micro;L||30&micro;L |- |NC||0.4&micro;L||0&micro;L||0&micro;L||1&micro;L||8.3&micro;L||10&micro;L |- |} {| class="wikitable" ! ||9/7 Pcon-RBS-tetR-DT||EcoRI||XbaI||SpeI||PstI||BufferD||BSA||MilliQ||total |- |2cut||14&micro;L||1&micro;L||1&micro;L||0&micro;L||0&micro;L||3&micro;L||18.2&micro;L||30&micro;L |- |NC||0.7&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.3&micro;L||10&micro;L |} {| class="wikitable" ! ||9/22 RBS-GFP-DT||EcoRI||XbaI||SpeI||PstI|| BufferD||BSA||MilliQ||total |- |2cuts||3.8&micro;L||1&micro;L||1&micro;L||0&micro;L||0&micro;L||3&micro;L||3&micro;L||18.2&micro;L||30&micro;L |- |NC||0.2&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.8&micro;L||10&micro;L |- |} {| class="wikitable" ! ||9/6 aptamer12-1R-DT||EcoRI||XbaI||SpeI||PstI||BufferD||BSA||MilliQ |- |2cut||8&micro;L||1&micro;L||1&micro;L||0&micro;L||0&micro;L||3&micro;L||3&micro;L||16&micro;L |- |NC||0.4&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.6&micro;L |} incubate 37&deg;C 1hour</div> ===Gel Extraction=== <div class="experiment"> <span class="author">No name</span> {| class="wikitable" !Lane||DNA||Enzyme |- |1||1kbp ladder||-- |- |3~5||Pcon-PT181 antisense (EcoRI&SpaI)||-- |} </div> ===Transformation=== <div class="experiment"> <span class="author">No name</span> {| class="wikitable" !Name||Sample||Competent Cells||Plate |- |9/20 Plac-aptamer12-1R-DT(XbaI&PstI)||2 &micro;L||20 &micro;L||Amp |- |9/20 pT181 attenuator(1C3)(EcoRI&SpeI)||2 &micro;L||20 &micro;L||CP |- |9/17 pT181 attenuator(1C3)(XbaI&PstI)||2 &micro;L||20 &micro;L||CP |- |9/20 Pcon-pT181 attenuator(SpaI&PstI)＋aptamer12-1R-DT(XbaI&PstI)||2 &micro;L||20 &micro;L||Amp |- |kaiABC||2 &micro;L||20 &micro;L||Amp |- |9/22 pSB1C3-RpaB(XbaI&PstI)||2 &micro;L||20 &micro;L||Amp |- |9/22 pSB1C3-pT181 attenuator(XbaI&PstI) ||2 &micro;L||20 &micro;L||Amp |- |9/22 Ptet(SpaI&PstI)＋RBS-GFP-DT(XbaI&PstI)||2 &micro;L||20 &micro;L||CP |- |9/22 RBS-GFP-DT＋Pcon-attenuator pT181 (EcoRI&SpeI)||2 &micro;L||20 &micro;L||CP |- |9/22 pT181 attenuator(XbaI&PstI)(1C3)||2 &micro;L||20 &micro;L||CP |} </div> ===PCR=== <div class="experiment"> <span class="author">No name</span> {| class="wikitable" !Sample||base pair |- |Pcon-RBS-tetR-DT(1-2Ⓐ)||-- |- |Pcon-RBS-tetR-DT(3-2Ⓐ)||-- |- |Pcon-RBS-GFP-DT(1-SⒶ)||-- |- |Pcon-RBS-GFP-DT(2-SⒶ)||-- |- |Pcon-tetR-DT(E-2Ⓐ)||-- |} {| class="wikitable" !PreDenature||Denature||Annealing||Extension||cycle |- |94 &deg;C||94 &deg;C||57 &deg;C||68 &deg;C||-- |- |2 min||10 s||30 s||1min5s||30 cycles |} </div> ===Electrophoresis=== <div class="experiment"> <span class="author">No name</span> {| class="wikitable" !Lane||Sample |- |1||1kbp ladder |- |2||Pcon-RBS-tetR-DT(1-2Ⓐ) |- |3||Pcon-RBS-tetR-DT(3-2Ⓐ) |- |4||Pcon-RBS-GFP-DT(1-SⒶ) |- |5||Pcon-RBS-GFP-DT(2-SⒶ) |- |6||Pcon-tetR-DT(E-2Ⓐ) |}

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