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Team:Tuebingen/Notebook/Protocols/mini-prep
From 2013.igem.org
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| - | + | Plasmid Preparation | |
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<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> | ||
| - | <li>On the next day, transfer culture to 15 mL Falcon tube and centrifuge for 10 min at 3500 rpm</li> | + | <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> | ||
| - | <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>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 | + | <li>In the end, elute with 30 µL Genaxxon Buffer 5.</li> |
| + | <li>Store at -20 °C.</li> | ||
</ol> | </ol> | ||
| + | |||
| + | <p> </p> | ||
| + | <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a> | ||
</div> | </div> | ||
Latest revision as of 23:36, 15 October 2013
Plasmid Preparation
Procedure
- 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.
- On the next day, transfer culture to 15 mL Falcon tube and centrifuge for 10 min at 3500 rpm.
- Discard supernatant completely.
- Resuspend cells in 250 µL Genaxxon Buffer 1.5 (with RNase)
- Transfer cells to Eppendorf-tube.
- Continue according to Genaxxon Plasmid DNA Purification Mini Prep Kit Manual.
- In the end, elute with 30 µL Genaxxon Buffer 5.
- Store at -20 °C.
Back to Protocols
