Team:Warsaw/Calendar-Main/28 May 2008

Rifampicin test #3 Michał L., Ewa, Marcin   Inoculation of liquid LB with E.coli TOP10 carrying pMPM-T5+AID 

 Inoculation of liquid LB with E.coli TOP10 carrying

pTrc99A+AID

Preparation of pMPMT5-AID+T7 and <a href=http://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPMT5+AID-T7</a> Michał K.   <a href="http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical <a href=http://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPM-T5 with transcriptional fusion</a>.</li>

 <a href="http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical <a href=http://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPM-T5 with translational fusion</a>.</li>

 Control <a href="http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of <a href=http://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPM-T5 with transcriptional fusion</a> with EcoRI (EcoRI buffer).</li>

 Control <a href="http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of <a href=http://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPM-T5 with translational fusion</a> with BamHI (BamHI buffer).</li>  Gel electrophoresis (<a href="http://2008.igem.org/Team:Warsaw/Calendar-Main/28_May_2008#fig1">Fig. 1</a> and <a href="http://2008.igem.org/Team:Warsaw/Calendar-Main/28_May_2008#fig2">Fig. 2</a>). </li>  Choice of proper clones and preparation for sequencing.</li> </ol>

<img src="http://2008.igem.org/wiki/images/6/62/Trawienie_06_05_2008.jpg" width=190/></a> Fig. 1. Here we found the clone with translational fusion on pMPM. These are different isolations in each lane. We have been working with the AT6 ever since. Digest was conducted with BamHI <img src="http://2008.igem.org/wiki/images/d/d6/Translation_fusion_digest_Eco.jpg"width=250/></a> Fig. 2. 2 to 6 - Control digests of few clones with transcriptional fusion; 7 - pMPMT5-AID; all plasmids digested with EcoRI.