Purification of proteins: A-alpha

Piotr

Transformation of Rosetta with pET15b+A-alpha plasmid. Plating on LB with chloramphenicol and ampicillin.

Blue-white screening and rifampicin test in GM2163

Michał L.

GM2163 competent cells carrying pZC320 were transformed with following plasmids:

• pMPMT5-omega,
• pMPMT5-AID,
• pMPMT5-AIDT7,
• pMPMT5-AID+T7.

Transformants were selected on CTAXI plates (LB + Chloramphenicol + Tetracycline + IPTG + Ampicillin 30 μg/mL + X-Gal) because GM2163 are chloramphenicol resistant.

Cloning of protein A DNA to pET15b+OmpA-alpha plasmid

Michał K.

1. Isolation of pET15b+A-alpha plasmids.
2. Control digest of isolated plasmids with XbaI and BamHI (Tango buffer).
3. Gel electrophoresis (Fig. 1.). Choice of proper clone.