Team:Warsaw/Calendar-Main/12 July 2008

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Preparation of construct pKS with A protein

Michał L., Ewa, Marcin

  1. Isolation of plasmid DNA from cultures inocluated on previous day.
  2. Control digest of isolated plasmid with SacI and NotI (BamHI buffer) - 2 h.
  3. Gel electrophoresis of digested DNA.
  4. We are glad to find the proper clone and inoculate liquid LB with ampicillin to freeze bacteria carrying pKS-A construct.

Cloning of protein Z DNA to pET15b-OmpA-alpha in place of OmpA

Piotr, Antoni

  1. Control digest of isolated plasmids with NdeI and NotI. Two positives obtained (proper band: ~200 bp visible).
  2. One of positive plasmids transformed into TOP10 and plated on LB+amp, overnight, for further isolation of pET15b+Z+alpha vector.

Preparation of constructs: OmpA_alpha and OmpA_omega #2

Michał K.

  1. Isolation of plasmids from cultures inocluated on previous day (pCACYC177 + OmpA_alpha).
  2. Control digest of isolated plasmids with BamHI and SacI (BamHI buffer).
  3. Gel electrophoresis - we found good clones (Fig. 1.).

Fig. 1. Control SacI/BamHI digests of plasmids which had a colony PCR product
1. Marker
2-4. digested plasmids which had a colony PCR product