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Protocol/4
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Revision as of 00:18, 26 October 2010
Protocol 4: Plasmid Miniprep
What you will need:
- Overnight culture
- Qiagen plasmid miniprep kit
Procedure:
- Transfer 1.5 ml of culture to Eppendorf.
- Spin down at 13 000 rpm.
- Discard supernatant and follow procedure on p. 22 of [http://www.qiagen.com/literature/render.aspx?id=370 QIAprep Miniprep Handbook].
- Determine concentration of purified plasmid with nanodrop
Notes
- Low speed pellet is easier to resuspend.
- To resuspend, take an empty micro centrifuge rack and rub the tube quickly over the holes, so it makes a "quacking" sound. It's pretty fun and works pretty good. Vortex after too.
- Step 2 in Qiagen, the alkaline lysis with buffer P2, should be brief to avoid damaging DNA.
