Preparation of constructs with OmpA protein fusions

1. Isolation of plasmids from cultures inocluated on previous day.
2. Control digest of isolated plasmids with BamHI and NotI (we confirmed pCACYC177 + OmpA_omega). We didn't obtain pCACYC177 + OmpA_alpha probably because of a mistake in plating.
3. Ligation of pACYC177 and OmpA_alpha (1 hr)
4. Transformation of E. coli TOP10 strain with ligation products: pACYC177 and OmpA_alpha.
5. Transformants plating on LB + kanamycin.

Preparation of construct pKS with A protein
Michał L., Marcin:

1. Inactivation of digestion enzymes and CIAP.
2. Ligation of digested PCR product and pKS 2h at room temperature.
3. Transformation of E. coli TOP10 strain with 7 µl of ligation mix.
4. Transformants plating on LB + ampicillin + X-gal + IPTG.