Team:Warsaw/Calendar-Main/4 June 2008

Preparation of pMPMT5-AID+AIDT7 construct Michał K.

Digest of DNA fragment (AIDT7) with HindIII and SalI (2x Tango buffer).

Digest of pMPM-T5 + AID with HindIII and SalI(2x Tango buffer), dephosphorylation (CIAP). Clean-up</a> of reaction products.</li> Ligation</a> of purified DNA (pMPM-T5 + AID with AID-T7 RNA-polymerase translation fusion).</li> Transformation</a> of E.coli TOP10</a> with ligation product.</li> Plating transformants on LB+tetracycline.</li></ol> Preparation of pMPMT5+T7 </a>construct Piotr Inoculation of separate transformant colonies (liquid LB + tetracycline).