Team:KULeuven/7 September 2009

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Project progress

Progress of parts

[edit] Blue Light Receptor

  1. Restriction digest with EcoRI and PstI on and the purified pcr product of in order to get our promotor on itself in the standard vector.
  2. Gel electroforesis and extraction of this RD. The concentration of this extraction was too low so no ligation was performed with these samples.
  3. Extra pcr was done on with primers 2260 and 2261.
  4. Liquid cultures of LigX and of were made from the 4x plates.

[edit] Vanillin Production

  • Miniprep of the four colonies from EF ligation
    • Accidentally used double volumes of elution buffer on EF4
part concentration 260/280 260/230
EF 1 115,5 1,98 2,31
EF 2 105,2 1,96 1,66
EF 3 95,1 2,05 0,98
EF 4 52,7 2,06 0,97
  • Gel electrophoresis from restriction on sam5 and sam8 looked good
  • Purified and ligated the restriction from sam5 and sam8 overnight
    • Concentration of sam8 was barely enough for ligation...
part concentration 260/280 260/230
sam5 10,1 2,11 0,04
sam8 3,1 3,96 0,02

[edit] Vanillin Receptor

  • a new restrictiondigest was done for puc and A because we decided that earlier results were not good enough.
  • There was a gelextraction, nanodrop and ligation done overnight.
  • The colony PCR of rpoA => 428 again for one minute => bad result
  • Miniprep of X+K was checked and put on gel. Wrong result probably because we selected a wrong colony.

[edit] Key/Lock/Anti-Key