Team:Tuebingen/Notebook/Protocols/mini-prep

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Mini-Prep
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Plasmid Preparation
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<a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a>
 
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<p>&nbsp;</p>
 
<h3>Procedure</h3>
<h3>Procedure</h3>
<ol>
<ol>
   <li>Inoculate 10 mL <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols/plates">LB-medium</a> with one single colony from a fresh plate (e.g. after <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols/trafo">transformation</a>) and incubate at 37 °C and 220 rpm over night.</li>
   <li>Inoculate 10 mL <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols/plates">LB-medium</a> with one single colony from a fresh plate (e.g. after <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols/trafo">transformation</a>) and incubate at 37 °C and 220 rpm over night.</li>
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   <li>On the next day, transfer culture to 15 mL Falcon tube and centrifuge for 10 min at 3500 rpm</li>
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   <li>On the next day, transfer culture to 15 mL Falcon tube and centrifuge for 10 min at 3500 rpm.</li>
   <li>Discard supernatant completely.</li>
   <li>Discard supernatant completely.</li>
   <li>Resuspend cells in 250 µL Genaxxon Buffer 1.5 (with RNase)</li>
   <li>Resuspend cells in 250 µL Genaxxon Buffer 1.5 (with RNase)</li>
   <li>Transfer cells to Eppendorf-tube.</li>
   <li>Transfer cells to Eppendorf-tube.</li>
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   <li>Continue according to <a href="http://www.genaxxon.de/Katalog/DNA-Reinigungskits/Plasmid-DNA-Purification-Mini-Prep-Kit-50-columns.html">Genaxxon Plasmid DNA Purification Mini Prep Kit Manual</a></li>
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   <li>Continue according to <a href="http://www.genaxxon.de/Katalog/DNA-Reinigungskits/Plasmid-DNA-Purification-Mini-Prep-Kit-50-columns.html">Genaxxon Plasmid DNA Purification Mini Prep Kit Manual</a>.</li>
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   <li>In the end, elute with30 µL Genaxxon Buffer 5.</li>
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   <li>In the end, elute with 30 µL Genaxxon Buffer 5.</li>
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  <li>Store at -20 °C.</li>
</ol>
</ol>
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<p>&nbsp;</p>
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<a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a>
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Latest revision as of 23:36, 15 October 2013

Return to iGEM Main Page.

Plasmid Preparation

Procedure

  1. Inoculate 10 mL LB-medium with one single colony from a fresh plate (e.g. after transformation) and incubate at 37 °C and 220 rpm over night.
  2. On the next day, transfer culture to 15 mL Falcon tube and centrifuge for 10 min at 3500 rpm.
  3. Discard supernatant completely.
  4. Resuspend cells in 250 µL Genaxxon Buffer 1.5 (with RNase)
  5. Transfer cells to Eppendorf-tube.
  6. Continue according to Genaxxon Plasmid DNA Purification Mini Prep Kit Manual.
  7. In the end, elute with 30 µL Genaxxon Buffer 5.
  8. Store at -20 °C.

 

Back to Protocols