Team:Warsaw/Calendar-Main/29 August 2008

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Purification of proteins: A-alpha

Piotr

Single transformations of Rosetta with plasmids pET15b+A-alpha. 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:

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.

Fig. 1. Control XbaI/BamHI digests of plasmids isolated from transformants with colony PCR product.
1. Marker
2-4. Digested plasmids pET15b+A-alpha



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