Cloning of protein A DNA to OmpA constructs

1. Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega and pACYC177+OmpA_A_alpha. Primers used: AL+SacI and AP+NotI
2. Confirmed transformant colonies (found only on pACYC177+OmpA_A_alpha plate) inoculated to liquid LB with kanamycin.

Paweł

1. Ligation transformed into TOP10 strain and plated on LB+kanamycin

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. Digest pET15b+OmpA_alpha and pGeneart+A with NdeI and NotI, pET15b+OmpA_alpha dephosphorylation with CIAP
3. Gel electrophoresis of digested plasmids and gel-out of proper bands
4. Overnight ligation of pET15b+alfa and A