Team:Warsaw/Calendar-Main/11 July 2008

Preparation of construct pKS with A protein Michał L., Ewa, Marcin   Colony PCR on colonies from plates with transformations pKS-A. Primers used:

AL+SacI and AP+NotI  Gel electrophoresis.  Confirmed transformant colonies inoculated to liquid LB with ampicillin.

Cloning of protein Z DNA to <a href=http://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b-OmpA-alpha</a> in place of OmpA Piotr, Antoni <a href=http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from cultures inoculated on previous day.

Preparation of constructs: OmpA_alpha and OmpA_omega #2

Michał K.   Colony <a href="http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> with <a href="http://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaL_N">OmpaL_N</a> and <a href="http://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaP_link">OmpaP_link</a>

primers on colonies from plates with transformations OmpA_alpha (annealing temperature - 55&deg;C,45 s of elongation step). </li>  Gel electrophoresis (<a href="http://2008.igem.org/Team:Warsaw/Calendar-Main/11_July_2008#fig1">Fig. 1.</a>).</li>  Confirmed transformant colonies inoculated to liquid LB with kanamycin.</li> </ol>

<img src="http://2008.igem.org/wiki/images/7/7e/11_th_july.jpg" width=300 /></a> Fig. 1. Colony PCR using: OmpaL_N and OmpaP_link primers Upper 1. Marker 2-13. colony PCR products carrying probable OmpA-alpha Lower 1. Marker 2-13. colony PCR products carrying probable OmpA-alpha