Team:Warsaw/Calendar-Main/12 July 2008

Preparation of construct pKS with A protein Michał L., Ewa, Marcin   Isolation of plasmid DNA from cultures inocluated on previous day.   Control digest of isolated plasmid with SacI and NotI (BamHI buffer) - 2 h.  Gel electrophoresis of digested DNA.  We are glad to find the proper clone and inoculate liquid LB with ampicillin to freeze bacteria carrying pKS-A construct.</li></ol>

Cloning of protein Z DNA to pET15b-OmpA-alpha</a> in place of OmpA Piotr, Antoni <ol> Control digest</a> of isolated plasmids with NdeI and NotI. Two positives obtained (proper band: ~200 bp visible).</li> One of positive plasmids transformed into TOP10</a> and plated on LB+amp, overnight, for further isolation of pET15b+Z+alpha</a> vector.</li> </ol> Preparation of constructs: OmpA_alpha and OmpA_omega #2 Michał K.  <ol>  Isolation of plasmids</a> from cultures inocluated on previous day (pCACYC177 + OmpA_alpha</a>). </li>  Control digest</a> of isolated plasmids with BamHI and SacI (BamHI buffer).</li>  Gel electrophoresis - we found good clones (Fig. 1.</a>).</li> </ol>

<img src="http://2008.igem.org/wiki/images/2/28/Cztery.jpg" width=300 /></a> Fig. 1. Control SacI/BamHI digests of plasmids which had a colony PCR product 1. Marker 2-4. digested plasmids which had a colony PCR product