Team:KULeuven/27 August 2009


Project progress

Progress of parts

[edit] Blue Light Receptor

  1. 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
  2. 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.
  3. 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

[edit] Key/Lock/Anti-Key