EPF-Lausanne/13 July 2009

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Contents


13 July 2009



Wet Lab

[http://partsregistry.org/Part:BBa_I6007 Inverter TetR (BBa_I6007)] was transformed again. And the new plasmid (created on July the 10th, 10.07.09) LacI-RBS was transformed on DH5-alpha competent cells.

We failed in purifing our [http://partsregistry.org/Part:BBa_B0010 Terminator (BBa_B0010)], do it again tomorrow, beginning with the digestion, etc.


Cloning Strategy

Restriction enzymes on [http://www.neb.com/nebecomm/products/category1.asp?#2 Biolabs website] and [http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_olignucleotides.asp clevage oligonucleotides]

TRP promoter biobrick strategy

  • Problem to overcome:
SpeI sites on Trp promoter sequence and it's an upstream part which has to be cut with ES.
  • Strategy:
PCR: Forward primer having E and X sites and Reverse primer NheI.
Digest Trp promoter with E and NheI.
Digest plasmid with E and X.
Ligation -> E site is recreated; X and NheI have compatible ends so ligation is possible and the site is destroyed (mixted site).


People in the lab

Heidi, Tu, Nath, Caro



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