Team:Warsaw/Calendar-Main/6 May 2008

Preparation of pMPMT5+AID construct Piotr, Weronika



Preparation of electro- and chemocompetent bacteria E. coli TOP10.

Setup of overnight cultures (pTrc99A-AID, pMPM-T5, pZC320) on LB broth with proper antibiotics.</li></ol>

Preparation of pMPMT5-AID+T7</a> and pMPMT5+AID-T7</a> Michał L.

 Isolation of genomic DNA</a> (E.coli Rosetta</a> and TOP10</a>) - template DNA for PCR to obtain T7 polymerase DNA. DNA isolated from E. coli TOP10 strain was used as a negative control for PCR. Fig. 1</a>.

<img src="http://2008.igem.org/wiki/images/3/3c/Genomic_DNA_WAW.jpg"/></a> Fig. 1. Probes of isolated genomic DNA after mechanical shearing by pipetting. Without shearing the DNA wouldn't be visible in this kind of gel because genomic DNA is too big. 1 and 3-E.coli Rosetta DNA; 2-DNA ladder; 4-E.coli TOP10 DNA.