PCR - AID for translational fusion.
Primers:
AIDlNrHAIDpLinB
Template DNA: pTrc99A-AID
Annealing temperature: 55 °C
Elongation time: 60 sec
20 cycles
PCR - T7 RNA-polymerase for translational fusion; Taq polymerase with supplemented buffer (Łukasz Banasiak helped us with that problematique PCR).
Primers:
T7lLinkBT7pXbSal
Template DNA: E. coliRosetta genomic DNA
Annealing temperature: 62 °C
Elongation time: 4 minutes
20 cycles
PCR - T7 RNA-polymerase for transcription fusion.
Primers:
T7lRBSHiT7pXbSal
Template DNA: E. coliRosetta genomic DNA
Annealing temperature: 62 °C
Elongation time: 4 minutes
20 cycles