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Template:Kyoto/Notebook/Sep 20
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==Sep 20== | ==Sep 20== | ||
| + | ===Ligation=== | ||
| + | <div class="experiment"> | ||
| + | <span class="author">Okazaki, Kanaoka, Tatsui and Hirano</span> | ||
| + | {| class="wikitable" | ||
| + | !state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2 | ||
| + | |- | ||
| + | |experiment||9/18 Plac (SpeI & PstI)||3.0 µL||9/18 RBS-lysis3-DT (XbaI & PstI)||12.4 µL||3.5 µL | ||
| + | |- | ||
| + | |experiment||9/12 Ptet-pT181 antisense||1.0 µL||9/18 spinach-DT||11.9 µL||3.5 µL | ||
| + | |- | ||
| + | |experiment||9/13 Pcon-RBS-luxR-DT||2.1 µL||9/10 Plux-RBS-GFP-DT||5.6 µL||3.5 µL | ||
| + | |- | ||
| + | |experiment||9/13 Pcon-pT181 attenuator (SpeI & PstI)||2.0 µL||9/18 aptamer 12-1R-DT (XbaI & PstI)||6.3 µL||3.5 µL | ||
| + | |- | ||
| + | |experiment||9/18 Plac ②(SpeI & PstI)||3.0 µL||9/18 aptamer 12-1R-DT(XbaI & PstI)||4.5µL||x3.5 µL | ||
| + | |- | ||
| + | |} | ||
| + | </div> | ||
| + | |||
| + | ===Transformation=== | ||
| + | <div class="experiment"> | ||
| + | <span class="author">Tatsui</span> | ||
| + | {| class="wikitable" | ||
| + | !Name||Sample||Competent Cells||Plate | ||
| + | |- | ||
| + | |9/17 pSB1C3-Pcon-pT181 attenuator-DT||2 µL||20 µL||CP | ||
| + | |- | ||
| + | |9/17 pSB1C3-Pcon-aptamer 12_1R-DT||2 µL||20 µL||CP | ||
| + | |- | ||
| + | |9/17 pSB1C3-Pcon-antisense-spinach-DT||2 µL||20 µL||CP | ||
| + | |- | ||
| + | |9/17 pSB1C3-Pcon-RBS-tetR-DT x2||2 µL||20 µL||CP | ||
| + | |- | ||
| + | |9/14 pSB1C3-PkaiBC||2 µL||20 µL||CP | ||
| + | |- | ||
| + | |9/14 pSB1C3-RpaB||2 µL||20 µL||CP | ||
| + | |- | ||
| + | |9/16 pSB1C3-pT181 attenuator(EcoRI&SpeI) ||2 µL||20 µL||CP | ||
| + | |- | ||
| + | |9/17 pSB1C3-pT181 attenuator(XbaI&PstI) ||2 µL||20 µL||CP | ||
| + | |- | ||
| + | |9/17 pSB1C3-Pcon-spinach-DT||2 µL||20 µL||CP | ||
| + | |- | ||
| + | |9/14 RBS-GFP-DT-PkaiBC||2 µL||20 µL||CP | ||
| + | |- | ||
| + | |Plux-RBS-lysis3-DT||2 µL||20 µL||CP | ||
| + | |- | ||
| + | |9/20 Ptet-pT181 antisense-spinach-DT||2 µL||20 µL||CP | ||
| + | |- | ||
| + | |9/20 Pcon-spinach-DT(pSB4K5) ||2 µL||20 µL||Kan | ||
| + | |- | ||
| + | |9/20 Pcon-pT181 antisense-spinach-DT(pSB4K5) ||2 µL||20 µL||Kan | ||
| + | |- | ||
| + | |9/20 KaiABC||2 µL||20 µL||Kan | ||
| + | |- | ||
| + | |9/20 KaiABC||2 µL||20 µL||Kan | ||
| + | |- | ||
| + | |9/20 Plac-RBS-lysis3-DT||2 µL||20 µL||Amp | ||
| + | |- | ||
| + | |9/20 Plac-aptamer 12_1R||2 µL||20 µL||Amp | ||
| + | |- | ||
| + | |9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT||2 µL||20 µL||Amp | ||
| + | |- | ||
| + | |Plac-aptamer12_1R-DT||2 µL||20 µL||Amp | ||
| + | |- | ||
| + | |9/20 Plac-aptamer12_1R-DT||2 µL||20 µL||Amp | ||
| + | |- | ||
| + | |9/20Pcon-pT181attenuator-aptamer-DT||2 µL||20 µL||Amp | ||
| + | |- | ||
| + | |9/17 Plac-pT181-attenuatorx2||2 µL||20 µL||Amp | ||
| + | |- | ||
| + | |9/15 Pcon-RBS-lacZα-DT||2 µL||20 µL||Amp | ||
| + | |- | ||
| + | |9/14 Pcon-pT181attenuator-RBS-lacZα-DT||2 µL||20 µL||Amp | ||
| + | |- | ||
| + | |PUC19||2 µL||20 µL||Amp | ||
| + | |} | ||
| + | </div> | ||
| + | |||
| + | ===PCR=== | ||
| + | <div class="experiment"> | ||
| + | <span class="author">Tatsui</span> | ||
| + | {| class="wikitable" | ||
| + | !Sample||base pair | ||
| + | |- | ||
| + | |pT181 attenuator(pSB1C3)||-- | ||
| + | |- | ||
| + | |Pcon-RBS-lacZα-DT||-- | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | !PreDenature||Denature||Annealing||Extension||cycle | ||
| + | |- | ||
| + | |94 °C||94 °C||55 °C||68 °C||-- | ||
| + | |- | ||
| + | |5 min||30 s||30 s||3 min||30 cycles | ||
| + | |} | ||
| + | </div> | ||
| + | |||
| + | ===Electrophoresis=== | ||
| + | <div class="experiment"> | ||
| + | <span class="author">No name</span> | ||
| + | {| class="wikitable" | ||
| + | !Lane||Sample||Enzyme | ||
| + | |- | ||
| + | |1||Pcon-RBS-lacZα-DT||-- | ||
| + | |- | ||
| + | |2||1kb ladder||-- | ||
| + | |- | ||
| + | |3||pSB1C3-pT181attenuator ||-- | ||
| + | |} | ||
| + | </div> | ||
Latest revision as of 11:39, 24 September 2013
Contents |
Sep 20
Ligation
| state | Vector | Inserter | Ligation High ver.2 | ||
|---|---|---|---|---|---|
| experiment | 9/18 Plac (SpeI & PstI) | 3.0 µL | 9/18 RBS-lysis3-DT (XbaI & PstI) | 12.4 µL | 3.5 µL |
| experiment | 9/12 Ptet-pT181 antisense | 1.0 µL | 9/18 spinach-DT | 11.9 µL | 3.5 µL |
| experiment | 9/13 Pcon-RBS-luxR-DT | 2.1 µL | 9/10 Plux-RBS-GFP-DT | 5.6 µL | 3.5 µL |
| experiment | 9/13 Pcon-pT181 attenuator (SpeI & PstI) | 2.0 µL | 9/18 aptamer 12-1R-DT (XbaI & PstI) | 6.3 µL | 3.5 µL |
| experiment | 9/18 Plac ②(SpeI & PstI) | 3.0 µL | 9/18 aptamer 12-1R-DT(XbaI & PstI) | 4.5µL | x3.5 µL |
Transformation
| Name | Sample | Competent Cells | Plate |
|---|---|---|---|
| 9/17 pSB1C3-Pcon-pT181 attenuator-DT | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-Pcon-aptamer 12_1R-DT | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-Pcon-antisense-spinach-DT | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-Pcon-RBS-tetR-DT x2 | 2 µL | 20 µL | CP |
| 9/14 pSB1C3-PkaiBC | 2 µL | 20 µL | CP |
| 9/14 pSB1C3-RpaB | 2 µL | 20 µL | CP |
| 9/16 pSB1C3-pT181 attenuator(EcoRI&SpeI) | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-pT181 attenuator(XbaI&PstI) | 2 µL | 20 µL | CP |
| 9/17 pSB1C3-Pcon-spinach-DT | 2 µL | 20 µL | CP |
| 9/14 RBS-GFP-DT-PkaiBC | 2 µL | 20 µL | CP |
| Plux-RBS-lysis3-DT | 2 µL | 20 µL | CP |
| 9/20 Ptet-pT181 antisense-spinach-DT | 2 µL | 20 µL | CP |
| 9/20 Pcon-spinach-DT(pSB4K5) | 2 µL | 20 µL | Kan |
| 9/20 Pcon-pT181 antisense-spinach-DT(pSB4K5) | 2 µL | 20 µL | Kan |
| 9/20 KaiABC | 2 µL | 20 µL | Kan |
| 9/20 KaiABC | 2 µL | 20 µL | Kan |
| 9/20 Plac-RBS-lysis3-DT | 2 µL | 20 µL | Amp |
| 9/20 Plac-aptamer 12_1R | 2 µL | 20 µL | Amp |
| 9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT | 2 µL | 20 µL | Amp |
| Plac-aptamer12_1R-DT | 2 µL | 20 µL | Amp |
| 9/20 Plac-aptamer12_1R-DT | 2 µL | 20 µL | Amp |
| 9/20Pcon-pT181attenuator-aptamer-DT | 2 µL | 20 µL | Amp |
| 9/17 Plac-pT181-attenuatorx2 | 2 µL | 20 µL | Amp |
| 9/15 Pcon-RBS-lacZα-DT | 2 µL | 20 µL | Amp |
| 9/14 Pcon-pT181attenuator-RBS-lacZα-DT | 2 µL | 20 µL | Amp |
| PUC19 | 2 µL | 20 µL | Amp |
PCR
| Sample | base pair |
|---|---|
| pT181 attenuator(pSB1C3) | -- |
| Pcon-RBS-lacZα-DT | -- |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94 °C | 94 °C | 55 °C | 68 °C | -- |
| 5 min | 30 s | 30 s | 3 min | 30 cycles |
Electrophoresis
| Lane | Sample | Enzyme |
|---|---|---|
| 1 | Pcon-RBS-lacZα-DT | -- |
| 2 | 1kb ladder | -- |
| 3 | pSB1C3-pT181attenuator | -- |
