Team:Tuebingen/Notebook/Protocols/transformation

From 2014.igem.org

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<h1>Protocols</h1>
<h3>Transformation of <i>E. coli</i></h3>
<h3>Transformation of <i>E. coli</i></h3>
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<h4>Procedure</h4>
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<ol>
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  <li>Add desired volume of DNA (not more than 1/10 volume, for transformation of ligations: add complete 10 μL ligation reaction) to the 100 μl-culture in microreaction-tube.</li>
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  <li>Incubate 30 min on ice.</li>
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  <li>Heat shock for 45 s at 42°C.</li>
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  <li>Incubate 10 min on ice.</li>
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 +
  <li>Add 0.9 ml of culture medium (no AB added) and let the bacteria grow at 37°C for 2h.</li>
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  <li>Plate on LB plates + antibiotic and incubate at 37°C o/n.</li>

Latest revision as of 15:29, 17 October 2014


Protocols

Transformation of E. coli

Procedure

  1. Add desired volume of DNA (not more than 1/10 volume, for transformation of ligations: add complete 10 μL ligation reaction) to the 100 μl-culture in microreaction-tube.
  2. Incubate 30 min on ice.
  3. Heat shock for 45 s at 42°C.
  4. Incubate 10 min on ice.
  5. Add 0.9 ml of culture medium (no AB added) and let the bacteria grow at 37°C for 2h.
  6. Plate on LB plates + antibiotic and incubate at 37°C o/n.

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