Team:Warsaw/Calendar-Main/4 July 2009
From 2009.igem.org
Cloning of p53 coding sequence
Marcin
Tasks:
- Transformation of chemocompetent E. coli strain DH5alpha
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°C
- incubation on the ice - 2 minuts
- add 800 ul of SOB medium to the bacteria
- incubation in 37°C - 1 h
- Plating on selective LB medium supplemented with Ampicilin, X-Gal and IPTG
Concluding remark:
Used bacteria was wrong but now we know where are correct bacteria.
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