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Team:DTU-Denmark/Notebook/20 August 2013
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| RV primer 55b || 3uL || 3uL || 3uL || 3uL || 3uL || 3uL || 3uL | | RV primer 55b || 3uL || 3uL || 3uL || 3uL || 3uL || 3uL || 3uL | ||
|- | |- | ||
| - | | DMSO 100% || 2.5uL (5%) || - || - || 2.5uL (5%) || - || - || - | + | | DMSO (100%) || 2.5uL (5%) || - || - || 2.5uL (5%) || - || - || - |
|- | |- | ||
| - | | Betaine 5M || - || 10uL (1M) || - || - || - || - || - | + | | Betaine (5M) || - || 10uL (1M) || - || - || - || - || - |
|- | |- | ||
|} | |} | ||
Revision as of 12:36, 20 August 2013
20 August 2013
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Contents |
lab 208
Main purpose
- PCR to linearize pZA21::RFP::araBAD
Who was in the lab
Kristian, Helen, Henrike, Julia
Procedure
PCR to linearize pZA21::RFP::araBAD
- Template: pZA21::RFP::araBAD
- Primers: 55a and 55b
- Program (A2 on the main machine)- Annealing temperature: 65C
The table shows the composition of each reaction:
| component (per reaction) | 1 | 2 | 3 | 4 | 5 | 6 | Neg |
|---|---|---|---|---|---|---|---|
| dNTPs | 1uL | 1uL | 1uL | 1uL | 1uL | 1uL | - |
| buffer (GC/HF + volume) | HF + 10uL | HF + 10uL | HF + 10uL | GC + 10uL | HF + 10uL | HF + 10uL | HF + 10uL |
| X7 polymerase | 0.5uL | 0.5uL | 0.5uL | 0.5uL | 0.5uL | 0.5uL | 0.5uL |
| MilliQ water | 29uL | 21.5uL | 31.5uL | 29uL | 31.5uL | 31.5uL | 32.5uL |
| template | 1uL | 1uL | 1uL | 1uL | 1uL | 1uL | - |
| FW primer 55a | 3uL | 3uL | 3uL | 3uL | 3uL | 3uL | 3uL |
| RV primer 55b | 3uL | 3uL | 3uL | 3uL | 3uL | 3uL | 3uL |
| DMSO (100%) | 2.5uL (5%) | - | - | 2.5uL (5%) | - | - | - |
| Betaine (5M) | - | 10uL (1M) | - | - | - | - | - |
Results
Gels
loaded more samples from the screening PCR for BioBrick creation yesterday
- 1 kb ladder
- screening D12
- screening D11
- screening D10
- screening D9
- screening D8
- screening D7
- screening D6
- screening D5
- screening D4
- screening D3
- screening D2
- screening D1
- screening C12
- screening C11
- screening C10
- screening C9
- screening C8
- 1 kb ladder
(pic big gel)
- 1 kb ladder
- screening C7
- screening C6
- screening C5
- screening C4
- screening C3
- screening C2
- screening C1
- screening B12
- screening B11
- screening B10
- screening B9
- screening B8
- screening B7
- 1 kb ladder
(pic small gel)
Conclusion
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