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DNA Construct Design and Cloning

 

Construct Design
  • The Facility can assist with the design of expression constructs. We use the Crystal Construct Designer (CCD) application to define optimal constructs in terms of solubility, domain boundaries etc., starting from the input DNA sequence. CCD combines multiple bio-informatic web-tools like secondary structure predictions, disorder predictions and domain definitions. CCD has been developed by Wijnand Mooij and Anastassis Perrakis and is available from the web.

 

LIC cloning
  • The facility has a variety of expression vectors, including a set of LIC (Ligation Independent Cloning) vectors for protein expression in E. coli, insect cells and mammalian cells. LIC cloning has many advantages over conventional cloning techniques: It is fast, does not require restriction enzymes (no incompatibility issues with internal cleavage sites) and the same PCR products can be inserted into different vectors.

  • Note: the actual cloning and construct preparation has to be done by the applicant!

NKI Protein Facility                                                                                                                                                                                                         Last updated: 17 April 2012

NKI-AvL

Plesmanlaan 121

1066 CX Amsterdam

The Netherlands