Team:Warsaw/Calendar-Main/19 May 2008

Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7 Michał K.  PCR - translational fusion: AID + T7 RNA-polymerase

Primers:

 AIDlNrH T7pXbSal</a></li> </ul>

Template DNA: purified PCR products from 16 May</a> - AID and T7 RNA-polymerase for translational fusion</a>

Annealing temperature : 73&deg;C Elongation time: 4 minutes

20 cycles </li>  Gel electrophoresis and isolation</a> of proper band (3300 bp). Fig. 1</a>.</li> Electrophoresis to estimate the concentration of isolated DNA.</li></ol>

<img src="http://2008.igem.org/wiki/images/e/e5/Pcr_fusion_WAW.jpg" width=190/></a> Fig. 1. 1-DNA ladder; 2-PCR product-translational fusion

Rifampicin test #1 Michał L., Ewa, Marcin

Overnight culture of the following strains: Top10</a> Top10</a> + 0.5 mM IPTG and Top10</a> carrying: pMPM-T5-AID</a> + Glc pTrc99a-AID</a> + Glc pMPM-T5-AID</a> + 0.01% L-Ara pTrc99a-AID</a> + 0.5 mM IPTG