Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7

Michał K.

1. PCR - AID for translational fusion.
   Primers: AIDlNrH AIDpLinB
   Template DNA: pTrc99A-AID
   Annealing temperature: 55°C
   Elongation time: 60 s
   20 cycles
2. PCR - T7 RNA-polymerase for translational fusion; Taq polymerase with supplemented buffer (Łukasz Banasiak helped us with that problematic PCR).
   Primers: T7lLinkB T7pXbSal
   Template DNA: E. coli Rosetta genomic DNA
   Annealing temperature: 62°C
   Elongation time: 4 minutes
   20 cycles
   
3. PCR - T7 RNA-polymerase for transcriptional fusion.
   Primers: T7lRBSHi T7pXbSal
   Template DNA: E. coli Rosetta genomic DNA
   Annealing temperature: 62°C
   Elongation time: 4 minutes
   20 cycles