Protocol/4

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Revision as of 00:33, 26 October 2010

TEAM ALBERTA

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.

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