Team:Warsaw/Calendar-Main/17 July 2008

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Cloning of protein A DNA to OmpA constructs

  1. Digest of pKS-A plasmid, pACYC177+OmpA_omega and pACYC177+OmpA_alpha with NotI and SacI (pACYC177 were also dephosporylated with CIAP)
  2. Gel electrophoresis and gel-out of proper bands (420 bp - pKS-A lane, 4050 bp - pACYC177+OmpA_omega lane and 4300 bp pACYC177+OmpA_alpha lane).
  3. Ligation of A DNA fragment with both pACYC177 vectors.
  4. Transformation of E. coli TOP10 strain with ligations.
  5. Transformants plating on LB + kanamycin.

Paweł: pET15b vector transformed into TOP10 strain for plasmid purification