Team:TU Darmstadt/protocols/Heat Shock Transformation

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<img alt="Problem" src="/wiki/images/6/66/Darmstadt_green_Problem.jpg" width="100" height="30"></a>
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<!-- Heat Shock Transformation  -->
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Revision as of 23:42, 4 October 2013





Heat Shock Transformation

Materials


Equipment

  • - Heatingbad
  • - Incubator


Chemicals & consumables

  • - Ice
  • - Pipets + steril tips
  • - dYT-Media
  • - LB-Agar-Plates + antibiotics


Procedure


  1. Defrost stocks of competent cells (100 µL in 1.5 ml Eppendorf tube) on ice.
  2. Add the DNA and incubate the suspension for 5 minutes on ice.
  3. Heat shock is done by incubating the cells for 1 minute at 42°C.
  4. Add 1 mL of DYT medium and incubate for 1 hour at 37°C in order to obtain antibiotic resistance.
  5. Finally the cells are ready to be crossed out.