Team:Alberta/Notebook/protocols/miniprep

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Revision as of 17:06, 26 October 2010

TEAM ALBERTA

Protocol 4: 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.


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