Team:Tuebingen/Notebook/Protocols/trafo

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<a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 16px">Back to Protocols</a>
 
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<h3>Procedure</h3>
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  <li>Add desired volume of (plasmid) DNA (not more than 1/10 vol.) to 100 µL <i>E. coli</i> culture (i.e. to <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols/chemo-competentcells">aliquot</a>) in Eppendorf-tube. </li>
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  <li>Incubate on ice for 30 min.</li>
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  <li>Heat shock at 42 °C for 90 s.</li>
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  <li>Incubate 10 min on ice.</li>
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  <li>Add 0.9 mL of <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols/lb">LB-medium</a> and let bacteria grow at 37°C.</li>
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  <li>Plate on <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols/plates">LB plates</a> + required amount of antibiotic and incubate at 37 °C over night.</li>
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  <li>If necessary, store plates at 4 °C.</li>
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<p><a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a></p>
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Latest revision as of 12:12, 4 October 2013

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E. coli Transformation

Procedure

  1. Add desired volume of (plasmid) DNA (not more than 1/10 vol.) to 100 µL E. coli culture (i.e. to aliquot) in Eppendorf-tube.
  2. Incubate on ice for 30 min.
  3. Heat shock at 42 °C for 90 s.
  4. Incubate 10 min on ice.
  5. Add 0.9 mL of LB-medium and let bacteria grow at 37°C.
  6. Plate on LB plates + required amount of antibiotic and incubate at 37 °C over night.
  7. If necessary, store plates at 4 °C.

Back to Protocols