Team:KULeuven/14 September 2009


Project progress

Progress of parts

[edit] Blue Light Receptor

  1. miniprep of ligY colonies. nannodrop: 111 ng/µl and 116 ng/µl.
  2. restriction digest of ligY miniprep
  3. gelelectrophoresis of restricted ligY and compared to ligA. the ligY fragment was larger than ligA which means that the ligation Y fauled. the promotor region probably ligated to RFP which was not restricted out of the vector properly.
  4. gel extraction of plasmid pSB1A2 (nanodrop: 10ng/µl)
  5. a new ligY was performed
  6. control of pcr-product . It was ok.

[edit] Vanillin Production

  • Miniprep of COMT
part concentration 260/280 260/230
COMT1 1 63,5 1,98
COMT1 2 99,4 1,95
COMT1 3 84,5 1,91
COMT2 1 92,1 2,00
COMT2 2 80,4 1,93
COMT2 3 74,2 1,95
COMT3 1 92,0 2,03
COMT3 2 62,2 2,04
COMT3 3 70,5 2,04
COMT4 1 93,4 1,91
COMT4 2 104,4 1,96
COMT4 3 81,9 2,01
  • Restriction with S/P, SAMS2 P (was already cut with E/X) and EF3 with E/S
  • Another attempt at electroporating EFT using EFT ligation from last week
  • Gel electroforesis of 5 μl from the restriction sample (30 μl)
    • Nothing in EF lane (no DNA???)
    • Promotor showed again two results of 900bp and 2k bp (wut is going on?)
  • An attempt was made to seperate the restriction enzymes by PCR purification on the remaining restriction sample, however nanodrop showed very confusing results, conclusion: let's not try this again. Next time heat inactivation of restriction enzymes to make sure they are inactive.
  • Made mother plate of SAMS

[edit] Vanillin Receptor

  • miniprep of virA in the uc vector, fragment was digested a the gelelectrophoresis showed a good result
  • new restriction digest for virB was done
  • TOPO-cloning for rpoA failed again, so due to deadlines we decided that there was to few time left to do the TOPO-cloning again or with new protocols and to do the mutagenesis also.

[edit] Key/Lock/Anti-Key