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

Antoni

1. Colony PCR on colonies from plates with transformations pGeneart+A Primers used: AP+NotI and AL+SacI.
2. Lack of confirmed transformant colonies.

Cloning of protein Z DNA to pET15b+Omp-alpha plasmid

Antoni

1. Digest pET15b+OmpA_alpha and pGeneart+Z with NdeI and NotI (Orange buffer), pET15b+OmpA_alpha dephosphorylation with CIAP.
2. Gel electrophoresis of digested plasmids and gel-out of proper bands (200 bp for Z lane and 6200 bp for pET15b lane).
3. Ligation of pET15b+alpha and Z.
4. Transformation of E. coli TOP10.
5. Transformants plating on LB + ampicillin.