10 mL SOC Medium
- 2 g Bacto-Tryptone
- 0.019 g KCl
- 0.5 g Bacto-Yeast extract
- 0.05 g NaCl
- 0.207 g MgCl2 · 6 H2O
- 0.036 g Glucose
were dissolved in 10 mL H2O and stored at 4°C in 1 mL aliquots.
Team:Hamburg/Notebook
100 mL TB PIPES Buffer
- 1.87 g KCl
- 0.17 g CaCl2
- 15.1 g PIPES
were dissolved in 100 mL H2O. NaOH was added to adjust the pH to 6.7.
Colony PCR I
PCR reaction Master Mix (20x)
| amount [μL] | |
| 10x DreamTaq Buffer | 40.0 |
| dNTPs, 10 mM | 8.0 |
| VF2 Primer, 10 μM | 20.0 |
| VR Primer, 10 μM | 20.0 |
| DreamTaq | 2.0 |
| H2O | 310.0 |
| 400.0 |
| Colony | cell with plasmids |
|---|---|
| 1 | NEB Competent Cells with pSB1A3/BBa_K081005/mTaz and pSB1C3/BBa_R0082/BBa_E0240 |
| 2 | NEB Competent Cells with pSB1A3/BBa_K081005/mTaz and pSB1C3/BBa_R0082/BBa_E0240 |
| 3 | NEB Competent Cells with pSB1A3/BBa_K081005/mTaz and pSB1C3/BBa_R0082/BBa_E0240 |
| 4 | NEB Competent Cells with pSB1A3/BBa_K081005/mTaz and pSB1C3/BBa_R0082/BBa_E0240 |
| 5 | JW 3367-3 with pSB1A3/BBa_K081005/mTaz and pSB1C3/BBa_R0082/BBa_E0240 |
| 6 | JW 3367-3 with pSB1A3/BBa_K081005/mTaz and pSB1C3/BBa_R0082/BBa_E0240 |
| 7 | JW 3367-3 with pSB1A3/BBa_K081005/mTaz and pSB1C3/BBa_R0082/BBa_E0240 |
| 8 | JW 3367-3 with pSB1A3/BBa_K081005/mTaz and pSB1C3/BBa_R0082/BBa_E0240 |
| 9 | JW 3367-3 with pSB1A3/BBa_K081005/mTaz and pSB1C3/BBa_R0082/BBa_E0240 |
| 10 | JW 3367-3 with pSB1A3/BBa_K608006/BBa_C0082 and pSB1C3/BBa_R0082/BBa_E0240 |
| 11 | JW 3367-3 with pSB1A3/BBa_K608006/BBa_C0082 and pSB1C3/BBa_R0082/BBa_E0240 |
| 12 | JW 3367-3 with pSB1A3/BBa_K608006/BBa_C0082 and pSB1C3/BBa_R0082/BBa_E0240 |
| 13 | JW 3367-3 with pSB1A3/BBa_K608006/BBa_C0082 and pSB1C3/BBa_R0082/BBa_E0240 |
| 14 | JW 3367-3 with pSB1A3/BBa_K608006/BBa_C0082 and pSB1C3/BBa_R0082/BBa_E0240 |
| 15 | JW 3367-3 with pSB1A3/BBa_K608006/BBa_C0082 |
Colonies were picked and resuspended in 4 μL H2O
3 μL H2O were transformed into 5 mL LB/CAmp/Amp-medium, the rest was used as template for the PCR.
Program: TAQSCREEN
result: all samples are positiv!
Funding
