Michał K:
1. PCR - translation fusion: AID + T7 RNA-polymerase - optymalization (temperature gradient 60°C - 80°C).
Primers:
AIDlNrH and
T7pXbSal
Template DNA: AID for translation fusion and T7 RNA-polymerase for translation fusion
Annealing time: 4 minutes
35 cycles
2. Gel electrophoresis.
3. Optimalization of PCR - translation fusion: AID + T7 RNA-polymerase - MgCl2 cocentration and number of cycles.
Primers:
AIDlNrH and
T7pXbSal
Template DNA: AID for translation fusion and T7 RNA-polymerase for translation fusion
Annealing time: 4 minutes
4. Gel electrophoresis.