Team:KULeuven/16 September 2009

From 2009.igem.org

Project progress

Progress of parts

[edit] Blue Light Receptor

  1. miniprep of pRB322 (nanodrop: 59,8ng/µl)
  2. ligY was replated
  3. logA electroporation
  4. 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.

[edit] Key/Lock/Anti-Key