Preparation of constructs with OmpA protein fusions
1. PCR on pBD (jaki numerek?) plasmid with
OmpaL_N and OmpaP_link
primers (15 cycles, annealing length 45s, annealing temperature 63 degrees)
2. PCR on pUC19 plasmid with
AlphaL_link and AlphaP_XB
primers (20 cycles, annealing length 45s, annealing temperature 63 degrees)
3. PCR on pUC19 plasmid with
OmegaL_link and OmegaP_EPB
primers (20 cycles, annealing length 30s, annealing temperature 58 degrees)
As a result we got three PCR products: OmpA_linker and two fragments of TEM1 beta-lactamese: linker_alpha and linker_omega.
4. Gel electrphoresis of PCR products and gel-out of proper bands