Team:Warsaw/Calendar-Main/18 June 2008

Preparation of constructs: OmpA_alpha and OmpA_omega Michał K. Isolation of pACYC177. Digest of purified pACYC177 with NdeI and BamHI (Tango 2x buffer).   Gel electrophoresis (Fig. 2) and isolation</a> of proper band (3200 bp).</li> Clean-up</a> of OmpA_alpha and OmpA_omega digest products (Fig. 1</a>). </li> Ligation</a> of purified DNA (pACYC177 with OmpA_alpha and OmpA_omega DNA fragments).</li> Transformation</a> of E.coli TOP10</a> with ligation products.</li> Plating transformants on LB+tetracycline.</li></ol>

<img src="http://2008.igem.org/wiki/images/c/c9/Ompa_Alpha_Omega_Cleanup_WAW.jpg" width=300/> </a> Fig. 1. OmpA_alpha (lane 2) and OmpA_omega (lane 3) digest products. <img src="http://2008.igem.org/wiki/images/5/5f/PACYC177_digest_WAW.jpg" width=300/></a> Fig. 2. pACYC177 digested with NdeI and BamHI (lane 2).

Blue/white and rifampicin test #2 Michał L., Ewa, Piotr AID in transcription fusion works like AID, but induce slower growth than AID; AID in translation fusion do not induce rifampicin resistance mutation