Team:Warsaw/Calendar-Main/5 July 2009

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(Difference between revisions)
(New page: {{WarNotebook}} <!-- do not edit above me! --><html> <h3>Cloning of p53 coding sequence</h3> <h4>Marcin</h4> <br/> <p>Tasks:</P> <ul> <li>Transformation of chemocompetent E. coli strain DH...)
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<li>add 800 ul of SOB medium to the bacteria</li>
<li>add 800 ul of SOB medium to the bacteria</li>
<li>incubation in 37&deg;C - 1 h</li>
<li>incubation in 37&deg;C - 1 h</li>
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<li>Plating on seleclive LB medium supplemented with Ampicilin, X-Gal and IPTG</li>
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<li>Plating on selective LB medium supplemented with Ampicilin, X-Gal and IPTG</li>
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Revision as of 00:04, 9 July 2009


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Cloning of p53 coding sequence

Marcin


Tasks:

  • Transformation of chemocompetent E. coli strain DH5alpha

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°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





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