From 2013.igem.org
Sep 20
Ligation
Okazaki,Kanaoka,Tatsui,Hirano
state | Vector | Inserter | Ligation High ver.2
|
experiment | 9/18 Plac(Spe1&Pst1) | 3.0µL | 9/18 RBS-lysis3-DT (XbaI&PstI) | 12.4µL | 3.5 µL
|
experiment | 9/12 Ptet-pT181antisense | 1.0µL | 9/18 spinach-DT | 11.9µL | 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-pT181attenuator(SpeI&PstI) | 2.0µL | 9/18 aptamer12-1R-DT (Xba1&Pst1) | 6.3µL | 3.5 µL
|
experiment | 9/18 Plac2(Spe1&Ps) | 3.0µL | 9/18 aptamer12-1R-DT(Xba1&Pst1) | 4.5µL | x3.5µL
|
Transformation
Tatsui
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
Kojima and Yoshida
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
No name
Lane | Sample | Enzyme
|
1 | Pcon-RBS-lacZα-DT | --
|
2 | 1kb ladder | --
|
3 | IC3-pT181attenuator | --
|