Team:Warsaw/Calendar-Main/25 September 2008

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Mutagenesis of protein A

Paweł

Treatment of mutageneses as on 23rd September.

Preparation of alpha_A construct

Antoni

  1. PCR on alpha_linker and linker_A with AlphaL+SacI and AP+NotI primers and 10% DMSO (30 cycles, elongation 60 s, annealing temperature 72°C).
  2. Gel electrophoresis of PCR products and gel-out of proper band (1019 bp).
  3. Measurment of concentration of both isolated products.

Preparation of BioBricks

Piotr

  1. Colony PCR with OmpaL_N and OmpaP_link primers on colonies from plates with transformations OmpA_alpha (45 s of elongation step).
  2. Plating on LB + ampicillin.