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Team:KULeuven/16 September 2009
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Revision as of 13:15, 19 September 2009 by JochemDeen (Talk | contribs)
Project progress
Progress of parts
[edit] Blue Light Receptor
- miniprep of pRB322 (nanodrop: 59,8ng/µl)
- ligY was replated
- logA electroporation
- plating of pSB3K3 from -80°C freezer.
[edit] Vanillin Production
- Restrictions from yesterday were put on gel (COMT2, 3, SAMS, Promotor)
- SAMS and promotor( J23110??) were gel purified
| part | concentration | 260/280 | 260/230 |
|---|---|---|---|
| SAMS | 8,5 | ||
| Promotor | 4,5 |
- Ligation of SAMS + promotor
- 28 μl of SAMS and 11 μl from promotor, almost 4:1 ratio (although maybe closer to 3:1)
- Electroporation of EFT ligation from yesterday on Km plates (Ter vector has Km resistance, so a closing of EF vector on its own doesn't grow!)
- Miniprep
| part | concentration | 260/280 | 260/230 |
|---|---|---|---|
| 1 | 69,2 | 1,99 | |
| 2 | 34,5 | 2,01 | |
| 3 | 74,9 | 2,03 |
- The liquid culture of SAMS was used for the glycerol stock and send for sequencing
[edit] Vanillin Receptor
- Gel-electrophoresis showed bad result for the mutation of virA probably due to too long elongation time. So we performed this PCR again with a new mutation.
- Ligation of virB didn't showed the expected result so we did it again.
