Griffitts:PCR clean-up

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Revision as of 12:56, 16 May 2011 by Matthew B. Crook (Talk | contribs)
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Materials

  • Buffer PB (in Qiagen kit)
  • Buffer PE (in Qiagen kit)
  • Buffer EB (in Qiagen kit)
  • QiaPrep spin column (1 per PCR reaction)
  • 1.5 mL microcentrifuge tube (1 per PCR reaction)

Procedure

  • Label spin columns and microcentrifuge tubes with the names of the appropriate PCR reactions
  • Using a P-200, add 200 μL buffer PB to each 40-μL PCR and mix well by gently pipetting up and down
  • Transfer the mixture to a QiaPrep spin column
  • Centrifuge at full speed for 30 seconds
  • Dump and tap
  • Add 700 μL buffer PE to each column
  • Centrifuge at full speed for 30 seconds
  • Dump and tap
  • Centrifuge for an additional 5 minutes at full speed
  • Move the column to a 1.5 mL microfuge tube
  • Add 50 μL buffer EB (to elute)
    • Drip this directly onto the filter disk but don't touch it with the pipette tip
  • Wait 2 minutes
  • Centrifuge at full speed for 1 minute
    • Point the lids of the microcentrifuge tubes in a clockwise direction, otherwise they'll break off
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