Preparation of constructs with OmpA protein fusions and Cloning of protein Z DNA to OmpA constructs
1. Isolation of plasmids from cultures inocluated on previous day (pCACYC177 + OmpA_alpha and pACYC177+OmpA_Z_omega).
2. Control digestation of isolated plasmids with BamHI and SacI (we found good clones for both ligations).
Linked PCR Omega-A
reisolated PCR product PCR-omega - 4 µl
reisolated PCR product A-homo - 13.5 µl
primer OmegaL+SacI_N - 2 µl
primer AP+NotI_N - 2 µl
Pfu buffer with Mg2+ - 5 µl
dNTPs - 1 µl
H2o - 22 µl

program:

1. 95°C - 3'
2. 95°C - 30"
3. 55°C - 45"
4. 68°C - 1'
5. go to step 2 25 x
6. 68° - 10'
7. keep in 4°

gel electrophoresis