Team:KULeuven/10 September 2009

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Revision as of 08:54, 10 September 2009 by Deepstar (Talk | contribs)

Project progress

Progress of parts

[edit] Blue Light Receptor

  1. Since the results from our light experiments keep failing, some new ideas were computed. pSB1A2 is the backbone for our construct but since this is a high copy number plasmid perhaps the ratio of promotor to repressor is unequal (not enough repressor). A new enting in LC was done with ligX. These were grown for a few hours at 37°C and then put in 16°C room overnight so that more repressor can be made (its promotor is temperature-sensitive).
  2. Ligation from 09/08 was electroporated in competent cells.
  3. Miniprep of LigX, restriction digest and gel electrophoresis. The signals were good so a motherplate for ligX was made. Also, the vector pSB1A2 was extracted from the gel (39,1 ng/µl).

Nanodrop results:

Part concentration (ng/μl) 260/280 λ
LigX1(1) 110,9 1,97
LigX1(2) 107,3 1,95
LigX1(3) 97,1 1,98
LigX1(4) 106,9 1,97
LigX2(1) 123,8 1,99
LigX2(2) 100,9 1,99
LigX2(3) 111 1,95
LigX2(4) 108 1,97

[edit] Vanillin Production

  • Gel electrophoresis of TER restriction
    • The vector for the TER is Psb1AK3 and has 3100bp, with an insert of ±3200 bp
    • Gel extraction
part concentration 260/280 260/230
TER 14,2 1,97
  • Created a motherplate from EF1
  • Ligated EF restriction + TER restriction
  • Transferred the SAMS 1 and SAMS 2 colonies to liquid medium

[edit] Vanillin Receptor

  • puc+A was plated
  • X+K was again elctroporated

[edit] Key/Lock/Anti-Key