Team:Warsaw/Calendar-Main/3 June 2008

Preparation of pMPMT5-AID+AIDT7 construct Michał K. DNA (PCR products from previous day) gel electrophoresis and isolation of DNA from proper band (20 cycles - 3300 bp). Fig. 1.

 Fig. 1. PCR results of translational fusion for the pMPMT5-AID+AIDT7 construct. Lane 1 - 25 cycles, lane 2 - 20 cycles.

Preparation of pMPMT5+T7 construct Piotr, Weronika



Digest of pMPMT5-AID+T7 (transcriptional fusion) with EcoRI and HindIII (2x Tango buffer).</li> <li>Use of T4 polymerase for  blunting</a> (3 hr of incubation).</li> <li>DNA gel electrophoresis.</li> <li>Isolation of DNA</a> from proper band (7000 bp).</li> <li>Ligation</a> of isolated DNA (pMPM-T5 + T7 RNA-polymerase</a>).</li> <li>Transformation</a> of E.coli TOP10</a> with ligation product.</li> <li>Plating transformants on LB + tetracycline.</li> </ol>