Team:Warsaw/Calendar-Main/5 July 2009

Cloning of p53 coding sequence Marcin Tasks:  Transformation of chemocompetent E. coli strain DH5&alpha; Comment: When we found have appropriate bacteria I transformed it using the ligation reaction prepared in 04.07.09 to create pKS plasmid with p53 coding sequence  Methods:  thaw bacteria on the ice - 10 minuts add 4 ul of ligation mixture to the bacteria incubation on the ice - 20 minutes heat shock - 1 minut, 42&deg;C incubation on the ice - 2 minuts add 800 ul of SOB medium to the bacteria</li> incubation in 37&deg;C - 1 h</li> Plating on selective LB medium supplemented with ampicillin, X-Gal and IPTG</li></ul>