Team:Warsaw/Calendar-Main/26 August 2008

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Cloning of protein A DNA to pET15b+Omp-alfa plasmid

Antoni

  1. Colony PCR on colonies from plates with transformations pGeneart+A Primers used: AP+NotI and AL+SacI
  2. Lack of confirmed transformant colonies

Cloning of protein Z DNA to pET15b+Omp-alfa plasmid

Antoni

  1. Digest pET15b+OmpA_alpha and pGeneart+A with NdeI and NotI, pET15b+OmpA_alpha dephosphorylation with CIAP
  2. Gel electrophoresis of digested plasmids and gel-out of proper bands
  3. Ligation of pET15b+alfa and A
  4. Transformation of E. coli TOP10.
  5. Transformants plating on LB + ampicillin.