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Template:Kyoto/Notebook/Sep 2
From 2013.igem.org
Contents |
Sep 2
Liquid Culture
| Sample | medium |
|---|---|
| 9/1 entA-(Master Plate)-1 | 4mL SOB(+Km) |
37°C
Colony PCR
| Sample | base pair |
|---|---|
| 9/1 RBS-lysis2-DT-(1) | 985 |
| Ptet-RBS-lacZα-DT-(1) | 765 |
| Plac-RBS-lacZα-DT-(1) | 765 |
| Plac-RBS-lacZα-DT-(2) | 765 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 1min | 30cycles |
Restriction Enzyme Digestion
| pSB1C3-(1) | EcoRI | SpeI | XbaI | PstI | BufferB | BufferD | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|---|
| 2 cuts(E+S) | 7µL | 1µL | 1µL | 0µL | 0µL | 3µL | 0µL | 0.3µL | 17.7µL | 30µL |
| NC(E+S) | 0.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0µL | 0.1µL | 8.6µL | 10µL |
| 2 cut(X+P)s | 7µL | 0µL | 0µL | 1µL | 1µL | 0µL | 3µL | 0.3µL | 17.7µL | 30µL |
| NC(X+P) | 0.3µL | 0µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0.1µL | 8.6µL | 10µL |
| 8/21 tRMA-spinach(1) | EcoRI | SpeI | BufferB | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 8µL | 1.0µL | 1.0µL | 3µL | 0.3µL | 17.6µL | 30µL |
| NC | 0.5µL | 0µL | 0µL | 1µL | 0.1µL | 8.4µL | 10µL |
| 8/21 tetR aptamer12_1R-(1) | EcoRI | SpeI | BufferB | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 6µL | 1.0µL | 1.0µL | 3µL | 0.3µL | 18.7µL | 30µL |
| NC | 0.4µL | 0µL | 0µL | 1µL | 0.1µL | 8.5µL | 10µL |
| 8/21 pT181 attenuator-(2) | EcoRI | SpeI | XbaI | PstI | BufferB | BufferD | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|---|
| 2 cuts(E+S) | 8µL | 1µL | 1µL | 0µL | 0µL | 3µL | 0µL | 0.3µL | 16.3µL | 30µL |
| NC(E+S) | 0.5µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0µL | 0.1µL | 8.4µL | 10µL |
| 2 cut(X+P)s | 8µL | 0µL | 0µL | 1µL | 1µL | 0µL | 3µL | 0.3µL | 16.3µL | 30µL |
| NC(X+P) | 0.5µL | 0µL | 0µL | 0µL | 0µL | 0µL | 1µL | 0.1µL | 8.4µL | 10µL |
Liquid Culture
Master Plate
Gel Extraction
EDTA Solution
Colony PCR
Master Plate
Liquid Culture
Colony PCR
