Team:KULeuven/12 August 2009

From 2009.igem.org

Project progress

Progress of parts

[edit] Blue Light Receptor

  1. Gel electrophoresis of BlP cut with EcoRI and SpeI
  2. Gel extraction of the Blp

Nanodrop results

Part concentration (ng/μl) 260/280 λ 260/230 λ
Blp 27 2,08
  1. PCR (with Pfx) of blue light promoter with primers iGEM 2260 and iGEM 2261
    • Annealing of 58C
  2. Inoculate liquid medium
  3. Ligation
Vector insert
pSB1A3+BBa_P1010 BBa_K238013 (BLP)
50ng -> 2,5 μl 5 ng -> 0,3 μl

[edit] Vanillin Production

  • Miniprep on sam5, sam8 and fcs
    • There was no growth on ech, thus new cultures in liquid medium
  • Restriction digest on sam5, sam8 and fcs
  • Gel electrophoresis on sam5, sam8 and fcs
  • Gel extraction on sam5 and fcs

Nanodrop results:

Part concentration (ng/μl) 260/280 λ 260/230 λ
fcs 4,2 6,08
sam5 7,3 2,76

Notes:

  • sam8 showed a very low concentration on the gel electrophoresis. Thus, it is better to redo this test.
  • 260/280 for fcs was very high and the concentration was not high enough. Therefore, we will redo the extraction and use multiple tubes on the same filter to increase the concentration

[edit] Vanillin Receptor

  • B,G and R were stored for further procedure. A failed again: there was no visible growth. This second time there was no problem with the competent cells because we used a commercial product instead of a self made. We assumed there was a problem with adding the polyA-tails to the PCR product because, when redoing the gelelectrophoresis, there were no errors with the PCR product.
  • We did a new PCR on the bacteria with the inserted plasmids for B,G and R to check if the gene was inserted correctly.
  • PCR was performed for W,X,Y and Z to isolate the genes.

[edit] Key/Lock/Anti-Key