Team:Tuebingen/Notebook/Protocols/mini-prep

From 2013.igem.org

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   <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>
   <li>In the end, elute with 30 µL Genaxxon Buffer 5.</li>
   <li>In the end, elute with 30 µL Genaxxon Buffer 5.</li>
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</ol>

Revision as of 02:12, 4 October 2013

Return to iGEM Main Page.

Mini-Prep
Back to Protocols

 

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.