From 2013.igem.org
Sep 20
Ligation
Okazaki, Kanaoka, Tatsui and Hirano
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
Tatsui
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
Tatsui
Sample | base pair
|
pT181attenuator(pSB1C3)(XbaI & PstI) | --
|
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
No name
Lane | Sample | Enzyme
|
1 | Pcon-RBS-lacZα-DT | --
|
2 | 1kb ladder | --
|
3 | IC3-pT181attenuator | --
|