PCRs for fusions

Michał K.:

1. Gel electrophoresis (15 May PCR's) and DNA isolation from proper bands (600 bp for AID lane and 2700 bp for T7 polymerase lanes).

2. PCR - translation fusion: AID + T7 RNA-polymerase - optimization (annealing temperature gradient 60°C - 80°C).

   Primers:

   AIDlNrH and T7pXbSal
   Template DNA: purified PCR products from 16 May - AID and T7 RNA-polymerase for translation fusion
   Elongation time: 4 minutes
   35 cycles
   

3. Optimization of PCR - translation fusion: AID + T7 RNA-polymerase - MgCl₂ cocentration and number of cycles.

   Primers:

   AIDlNrH and T7pXbSal

   Template DNA: purified PCR products from 16 May - AID and T7 RNA-polymerase for translation fusion
   Annealing temperature: 73°C
   Elongation time: 4 minutes
   
4. Gel electrophoresis of PCR products.