Team:KULeuven/27 August 2009
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Revision as of 11:05, 27 August 2009
Project progress
Progress of parts
[edit] Blue Light Receptor
- The electroporations of 26/08 were checked.
- had some colonies. they were ented in liquid culture.
- LigC ( + ) did not grow. we figured that an insert of 35bp was too short to ligate so we decided to use as insert and + as vector . The following restriction digest was started:
- was cut with SpeI and PstI
- was cut with XbaI and PstI
- The miniprep that was made to sequence the LigA construct on 26/08 was used again to perform a restriction digest (EcoRI and PstI). This was put on gel to check whether there actually was LigA-insert in the vector and whether the insert had the right length. The gel showed a signal at 1000 bp and at 2000 bp which coincides with the insert (BLp + GFP) and the vector.
- A new setup to light the E.coli was engineered.
- Fresh cultures were made from the old ones (LB plate ligA 14/08 and the two liquid cultures from 26/08 were used as templates.)
- They were put in the 16°C room for about an hour
- A blue light (40W) was put on them for about an hour
- They were put in the 37°C incubator overnight
[edit] Vanillin Production
- Restriction4Real: we performed a gel electrophoresis together with opened plasmids (cut with EcoRI) to compare sizes
Part | DNA µl | MQ µl |
---|---|---|
Sam8 B | 7,9 | 18,1 |
Sam5 B | 4,0 | 22,0 |
Ech C | 4,75 | 21,25 |
Fcs B | 4,4 | 21,6 |
- Gel signals were inconclusive: gel ran not long enough
[edit] Vanillin Receptor
- Because of bad results yesterday the transformation of W was extra checked by performing a new PCR with different primers :
sample A : M13forward - virAforward sample B : M13forward - virAreverse sample C : virAforward - virAreverse sample D : M13reverse - virAforward sample E : M13reverse - virAreverse
Afterwards was gelelectrophoresis done and this showed virA was not present in the cell.
- ligation of X and Y showed a pover result so new ligations were done for virB and virG