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Template:Kyoto/Notebook/Sep 20
From 2013.igem.org
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==Sep 20== | ==Sep 20== | ||
===Ligation=== | ===Ligation=== | ||
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<div class="experiment"> | <div class="experiment"> | ||
| - | <span class="author">Okazaki,Kanaoka,Tatsui | + | <span class="author">Okazaki, Kanaoka, Tatsui and Hirano</span> |
{| class="wikitable" | {| class="wikitable" | ||
!state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2 | !state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2 | ||
|- | |- | ||
| - | |experiment||9/18 Plac(SpeI&PstI)||3.0µL||9/18 RBS-lysis3-DT (XbaI&PstI)||12.4µL||3.5 µL | + | |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- | + | |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||2.1µL||9/10 Plux-RBS-GFP-DT||5.6µL||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- | + | |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 | + | |experiment||9/18 Plac ②(SpeI & PstI)||3.0 µL||9/18 aptamer 12-1R-DT(XbaI & PstI)||4.5µL||x3.5 µL |
|- | |- | ||
|} | |} | ||
</div> | </div> | ||
| - | |||
===Transformation=== | ===Transformation=== | ||
Revision as of 09:36, 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 IC3-Pcon-pT181attenuator-DT | 2µL | 20µL | CP |
| 9/17 IC3-Pcon-aptamer12-1R-DT | 2µL | 20µL | CP |
| 9/17 IC3-Pcon-antisense-spinach-DT | 2µL | 20µL | CP |
| 9/17 IC3-Pcon-RBS-tetR-DTx2 | 2µL | 20µL | CP |
| 9/14 IC3-PkaiBC | 2µL | 20µL | CP |
| 9/14 IC3-RpaB | 2µL | 20µL | CP |
| 9/16 IC3-pT181attenuator(EcoRI&SpeI) | 2µL | 20µL | CP |
| 9/17 IC3-pT181attenuator(XbaI&PstI) | 2µL | 20µL | CP |
| 9/17IC3-Pcon-spinach-DT | 2µL | 20µL | CP |
| 9/14 RBS-GFP-DT-PkaiBC | 2µL | 20µL | CP |
| Plux-RBS-lysis3-DT | 2µL | 20µL | CP |
| 9/20 Ptet-pT181antisense-spinach-DT | 2µL | 20µL | CP |
| 9/20 Pcon-spinach-DT(4K5) | 2µL | 20µL | K |
| 9/20 Pcon-pT181antisense-spinach-DT(4K5) | 2µL | 20µL | K |
| 9/20 KaiABC | 2µL | 20µL | K |
| 9/20 KaiABC | 2µL | 20µL | K |
| 9/20 Plac-RBS-lysis3-DT | 2µL | 20µL | Amp |
| 9/20 Plac-aptamer12-1R | 2µL | 20µL | Amp |
| 9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT | 2µL | 20µL | Amp |
| Plac-aptamer12-1R-DT | 2µL | 20µL | Amp |
| 9/20 Plac-aptamer12-1R-DT | 2µL | 20µL | Amp |
| 9/20Pcon-pT181attenuator-aptamer-DT | 2µL | 20µL | Amp |
| 9/17 Plac-pT181-attenuatorx2 | 2µL | 20µL | Amp |
| 9/15 Pcon-RBS-lacZα-DT | 2µL | 20µL | Amp |
| 9/14 Pcon-pT181attenuator-RBS-lacZα-DT | 2µL | 20µL | Amp |
| PUC19 | 2µL | 20µL | Amp |
PCR
| Sample | base pair |
|---|---|
| pT181attenuator(IC3)(XbaI&PstI) | -- |
| Pcon-RBS-lacZα-DT | -- |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 3min | 30cycles |
Electrophoresis
| Lane | Sample | Enzyme |
|---|---|---|
| 1 | Pcon-RBS-lacZα-DT | -- |
| 2 | 1kb ladder | -- |
| 3 | IC3-pT181attenuator | -- |
