Difference between revisions of "Team:Birkbeck/Chemical Transformation"

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<h1>Chemical Transformation</h1>
 
<h1>Chemical Transformation</h1>
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<p><b>See previous protocol: <a href="https://2015.igem.org/Team:Birkbeck/Chemical_Competent_E._coli_Cell_Protocol">Chemical Competent <i>E. coli</i> Cell Protocol</b> </a>for details on making competent <i>E. coli</i> culls.</p>
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<p><b>See previous protocol: <a href="https://2015.igem.org/Team:Birkbeck/Chemical_Competent_E._coli_Cell_Protocol">Chemical Competent <i>E. coli</i> Cell Protocol</b> </a>for details on making chemically competent <i>E. coli</i> cells.</p>
 
<h2><b>Chemical Transformation protocol.</b></h2>
 
<h2><b>Chemical Transformation protocol.</b></h2>
 
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Revision as of 18:24, 11 September 2015

Chemical Transformation

See previous protocol: Chemical Competent E. coli Cell Protocol for details on making chemically competent E. coli cells.

Chemical Transformation protocol.

  • 1. Thaw out the 50 µL cell suspension aliquot on ice.
  • 2. Add 100pg-1µg of plasmid DNA to the cell suspension (approximately 1-5 µL of plasmid solution).
  • 3. Mix plasmid and cells thoroughly and incubate on ice for 30 minutes.
  • 4. Induce heat shock by incubating cells at 42oC for 30 seconds.
  • 5. Incubate on ice for 5 minutes.
  • 6. Add 450 µL of LB and incubate at 37oC for 1 hour.
  • 7. Plate out 50 µL of straight transformant cells & also carry out a 10-fold dilution (plate out 50µL). Note that the plate should contain the appropriate antibiotic at the MBC in order to select for transformants which contain the desired plasmid.
  • 8. Incubate overnnight at 37oC in a static incubator.