Team:Warsaw/Calendar-Main/7 July 2008

Preparation of constructs: OmpA_alpha and OmpA_omega #2 Piotr, Weronika It's the second attempt to clone OmpA_alpha and OmpA_omega to pACYC177, but now thank's to Paweł we've got both fusions on pET15b.   Digest of confirmed clones of pET15b+OmpA_alpha and pET15b+OmpA_omega with NdeI and BamHI (Tango 2x buffer).   Digest of pACYC177 plasmid with NdeI and BamHI (Tango 2x buffer), dephosphorylation with CIAP. </li>  Gel electrophoresis of digested plasmids and gel-out</a> of proper bands (OmpA_alpha- 1000 bp, OmpA_omega - 800 bp). </li> Electrophoresis of gel-out products.</li>  Overnight ligation</a> of pACYC177</a> and OmpA_alpha. </li>  Overnight ligation</a> of pACYC177</a> and OmpA_omega. </li> </ol>

Cloning omega-A fusion on pKS (second attempt) Michał L., Ewa, Marcin We have just received corrected version of omegaP-link10-homo2 primer. Let's hope this time everything will be fine. We are repeating the PCRs: