Preparation of constructs with OmpA protein fusions

Michał K.

1. Isolation of plasmids from cultures inocluated on previous day (pCACYC177 + OmpA_alpha).
2. Control digest of isolated plasmids with BamHI and SacI (BamHI buffer).
3. Gel electrophoresis - we found good clones.

Preparation of construct pKS with A protein

Michał L., Ewa, Marcin

1. Isolation of plasmid DNA from cultures inocluated on previous day.
2. Control digest of isolated plasmid with SacI and NotI (BamHI buffer) - 2 h.
3. Gel electrophoresis of digested DNA (Fig. 1.).
4. We are glad to find the proper clone and inoculate liquid LB with ampicillin to freeze bacteria carrying pKS-A construct.

Cloning of protein Z DNA to pET15b-OmpA-alpha in place of OmpA

Piotr, Antoni

1. Control digest of isolated plasmids with NdeI and NotI. Two positives obtained (proper band: ~200 bp visible).
2. One of positive plasmids transformed into TOP10 and plated on LB+amp, overnight, for further isolation of pET15b+Z+omega vector.