Cloning of protein A DNA to pET15b+Omp-alfa plasmid

Antoni

1. Isolation of plasmids from cultures inocluated on previous day (pGeneart+A and pET15b+OmpA-alfa).
2. Control digest of pGeneart+A plasmids with NdeI and NotI (Orange buffer).
3. Gel elctrophoresis. Chioce of proper clone.
4. Digest pET15b+OmpA_alpha and pGeneart+A with NdeI and NotI (Orange buffer), pET15b+OmpA_alpha dephosphorylation with CIAP.
5. Gel electrophoresis of digested plasmids and gel-out of proper bands (400 bp for protein A lane and 6200 for pET15b lane).
6. Overnight ligation of pET15b+alfa and A.