Preparation of constructs with OmpA protein fusions
Michał K.

1. PCR on OmpA_linker and linker_alpha with OmpaL_N and AlphaP_XB primers (20 cycles, elongation length 1 min 15 s, annealing temperature 57°C).
2. PCR on OmpA_linker and linker_omega with OmpaL_N and OmegaP_EPB primers (20 cycles, elongation length 1 min 15 s, annealing temperature 57°C).
3. Gel electrophoresis of PCR products and gel-out of OmpA_alpha and OmpA_omega.
4. Overnight digest of purified OmpA_alpha and OmpA_omega DNA with NdeI and BamHI.
5. Overnight digest of pET15b plasmid with NdeI and BamHI, dephosphorylation with CIAP.