Team:Warsaw/Calendar-Main/4 July 2009

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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°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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