Team:Warsaw/Calendar-Main/15 October 2009

Creation of the cro-box testing devices Kamil

Tasks:  Reporter device carrying plasmid digestion Cro-box carrying plasmids digestion 

Methods:  For the reporter device the digest mix was prepared as follows: 10&mu;l purified plasmid 2&mu;l Tango Buffer (Fermentas) 1&mu;l XbaI Enzyme (Fermentas) 1&mu;l PstI Enzyme (Fermentas) 5&mu;l H2O  For the cro-box the digest mix was prepared as follows (same for both plasmids): 10&mu;l purified plasmid 2&mu;l Tango Buffer (Fermentas) 1&mu;l BcuI Enzyme (Fermentas) 1&mu;l PstI Enzyme (Fermentas) 5&mu;l H2O  The digest was carried out in 37&deg;C for 4h. The results were visualized on 1% agarose gel. Gel extraction and purification of the appropriate bands was carried out using the Gel-Out kit (A&A Biotechnology) </ul>

Mutagenesis of cI Kuba

Task: Obtain a functional, termosensitive version of cI


 * inactivation ligation (15', 65oC)
 * Dpn1 digest (5 h)
 * transformation into E. coli (Top10)