Team:Warsaw/Calendar-Main/4 July 2009

Cloning of p53 coding sequence Marcin Tasks:  Transformation of chemocompetent E. coli strain DH5alpha  Comment: We was not sure whether the chosen bacteria had been working and we transformed the bacteria with pKS plasmid Methods:  thaw bacteria on the ice - 10 minuts add 1 ul of empty pKS plasmid 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 Ampicilin, X-Gal and IPTG</li></ul> Concluding remark: Used bacteria was wrong but now we know where are correct bacteria.