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Team:DTU-Denmark/Notebook/29 August 2013
From 2013.igem.org
29 August 2013
Contents |
lab 208
Main purpose
Who was in the lab
Procedure
===Introduce endings for USER cloning in pSB1C3 The table shows the composition of each reaction:
| component | 1 | 2 | 3 | Neg |
|---|---|---|---|---|
| dNTPs | 1uL | 1uL | 1uL | 1uL |
| HF buffer | 10uL | 10uL | 10uL | 10uL |
| X7 polymerase | 0.5uL | 0.5uL | 0.5uL | 0.5uL |
| MilliQ water | 28.5uL | 28.5uL | 28.5uL | 29.5uL |
| template pSB1C3 | 1uL | 1uL | 1uL | - |
| FW primer 57a2 | 3uL | 3uL | 3uL | 3uL |
| RV primer 57b | 3uL | 3uL | 3uL | 3uL |
| DMSO (100%) | 2uL (4%) | 2uL (4%) | 2uL (4%) | 2uL (4%) |
| MgCl2 (50mM) | 1uL (1mM) | 1uL (1mM) | 1uL (1mM) | 1uL (1mM) |
Results
Gel pictures
Ran gels on yesterday's PCRs to create psB1C3 with USER endings.
Small gel:
- 1 kb ladder
- 2%, pur
- 4%, pur
- 5%, pur
- 4%, 1uL, pur
- 4%, 2uL, pur
- 1M, pur
- 2%, HQ
- 4%, HQ
- 5%, HQ
- 4%, 1uL, HQ
- 4%, 2uL, HQ
- 1M HQ
- 1 kb ladder (ladder was almost empty so I put another one in the next lane)
- 1 kb ladder
big gel (samples from touch down PCR):
- 1 kb ladder
- GC, HQ
- 2% HQ
- 4% HQ
- 5% HQ
- 4%, 1uL, HQ
- 4%, 2uL, HQ
- 1M HQ
- GC pur
- 2%, pur
- 4%, pur
- 5%, pur
- 4%, 1uL, pur
- 4%, 2uL, pur
- 1M pur
- neg
- 1 kb ladder
Conclusion
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