Team:Alberta/Notebook/protocols/miniprep
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Latest revision as of 02:34, 27 October 2010
Plasmid Miniprep
Reagents:
- Overnight culture
- Qiagen plasmid miniprep kit
Procedure:
- Transfer 1.5ml 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 apparatus and vortex.
- Step 2 in Qiagen, the alkaline lysis with buffer P2, should be brief to avoid damaging DNA.