Preparation of constructs with OmpA protein fusions

Piotr:

1. Digest of confirmed clones of pET15b+OmpA_alpha and pET15b+OmpA_omega with NdeI and BamHI.
2. Digest of pACYC177 plasmid with NdeI and BamHI, dephosphorylation with CIAP.
3. Gel electrophoresis of digested plasmids and gel-out of proper bands (OmpA_alpha- 1000 bp, OmpA_omega - 800 bp).
4. Electrophoresis of gel-out products.
5. Overnight ligation of pACYC177 and OmpA_alpha.
6. Overnight ligation of pACYC177 and OmpA_omega.

Cloning omega-A fusion on pKS (second attempt)

Michał L., Ewa, Marcin:

We have just received corrected version of omegaP-link10-homo2 primer. Let's hope this time everything will be fine. We are repeating the PCRs:

PCR program for linker-A and omega-linker