Team:KULeuven/13 August 2009

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Revision as of 15:24, 16 August 2009

Project progress

Progress of parts

[edit] Blue Light Receptor

  1. PCR product of 12-august of pairt
  2. Miniprep of

Nanodrop results:

Part concentration (ng/μl) 260/280 λ 260/230 λ
123,3 1,82
141,2 1,99
  1. Restriction digest cut with EcoRI and XbaI
  2. Electroporation of + and the ligation of ...
  3. Gelelectrophoresis of
    • Only 1 band, so probably everything was cut
  4. Gelextraction of
    • Nanodrop = 29,3 ng/μl
  5. Ligation of vector and insert

[edit] Vanillin Production

  1. Plated COMT (+RBS) from the registry
    • Growing overnight in 37°C
  2. Plated ech
    • From newly made liquid culture
  3. Miniprep of ech and sam8
    • Nanodrop results:
Part concentration (ng/μl) 260/280 λ 260/230 λ
ech 72,7 1,94
sam8 66,2 2,00
  1. Restriction digest on ech and sam8
  2. Gel electrophoresis on ech and sam8
  3. New electrocompetent cells were used for the electroporation with the old ligations
    • Lig A -> sam5 + sam8
    • Lig C -> ech + fcs
    • Note: there was no spark
  4. Plating of the electroporated cells
    • 200 μl per plate twice
    • 600 μl left of A and C

[edit] Vanillin Receptor

  1. B and R miniprepped. Tomorrow proceed with mutagenesis of RpoA
  2. G redone (only TOPO was visible, 200bp) PCR from 2,3 and 1'
  3. W, X, Y, Z PCR purification to create a TOPO vector tomorrow

Nanodrop:

Part concentration (ng/μl) 260/280 λ 260/230 λ
W 46,1 1,84
X 112 1,82
Y 207 1,85
Z 155,3 1,86

[edit] Key/Lock/Anti-Key