Team:Warsaw/Calendar-Main/11 July 2008

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Preparation of construct pKS with A protein

Michał L., Ewa, Marcin

  1. Colony PCR on colonies from plates with transformations pKS-A.
    Primers used: AL+SacI and AP+NotI
  2. Gel electrophoresis.
  3. Confirmed transformant colonies inoculated to liquid LB with ampicillin.

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

Piotr, Antoni

Isolation of plasmids from cultures inoculated on previous day.

Preparation of constructs: OmpA_alpha and OmpA_omega #2

Michał K.

  1. Colony PCR with OmpaL_N and OmpaP_link primers on colonies from plates with transformations OmpA_alpha (annealing temperature - 55°C,45 s of elongation step).
  2. Gel electrophoresis (Fig. 1.).
  3. Confirmed transformant colonies inoculated to liquid LB with kanamycin.

Fig. 1. Colony PCR using: OmpaL_N and OmpaP_link primers
Upper
1. Marker
2-13. colony PCR products carrying probable OmpA-alpha
Lower
1. Marker
2-13. colony PCR products carrying probable OmpA-alpha