Mutagenesis of protein A

Paweł

1. Results of mutagenesis: no colonies on any plate.
2. Mutagenesis repeated with modified conditions: fresh stock of dNTPs used and additional MgCl2 added.

Preparation of alpha+A construct

Antoni

1. PCR on pKS plasmid containing protein A with AL+link10+homo2 and AP+NotI primers (20 cycles, elongation duration 15 s, annealing temperature 72°C).
2. PCR on pUC19 plasmid with AlphaL+SacI and AlphaP+link10+homo2 primers (20 cycles, elongation duration 45 s, annealing temperature 63°C).
   As a result we got two PCR products (alpha-linker and linker-A) wich will be utilized as templates in next PCR.
3. Gel electrophoresis of PCR products and gel-out of proper bands (alpha_linker - 572 bp and linker_A - 467 bp ).
4. Measurment of concentration of both isolated products