Team:Warsaw/Calendar-Main/22 May 2008

Rifampicin test #2 Piotr

Overnight inoculation and induction: Top10

and Top10 carrying: pMPM-T5-AID + 0.01% L-Ara pTrc99a-AID + 0.5 mM IPTG pMPM-T5-AID pTrc99a-AID

Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7 Piotr, Michał K. <ol><li> Design of primers for sequencing something we want from pMPM-T5.

Primers:

<ul> <li> pMPMT5seqF</a></li> <li>pMPMT5seqR</a></li> <li>AIDseqP</a></li></ul>

<li> Isolation</a> of hypothetical pMPM-T5 with transcriptional fusion</a>.</li>

<li> Isolation</a> of hypothetical pMPM-T5 with translational fusion</a..</li>

<li> Control digest</a> of pMPM-T5 with transcriptional fusion</a> with EcoRI (EcoRI buffer).</li>

<li> Control digest</a> of pMPM-T5 with translational fusion</a> with BamHI (BamHI buffer).</li>

<li> Gel electrophoresis (no proper colonies found). Fig. 1</a>.</li> <img src="http://2008.igem.org/wiki/images/8/87/Aid_fusion_digests_WAW.jpg"width=300/></a> Fig. 1. 1-DNA ladder; 2 and 3-hypothetical pMPM-T5 with transcriptional fusion digested with EcoRI (2 clones); 4-pMPM-T5-AID digested with EcoRI; 5-hypothetical pMPM with translational fusion digested with BamHI; 6-pMPM-T5-AID digested with BamHI.