Team:Newcastle/PCR purification


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PCR Purification


  • 1.5 ml microcentrifuge tubes
  • 2 ml collection tube
  • QIAquick columns
  • Qiagen Buffer PB
  • Qiagen Buffer EB
  • Qiagen Buffer PE
  • DNA mixture from PCR


  1. Add 5 volumes of Qiagen Buffer PB to 1 volume of PCR product.
  2. Put a QIAquick spin column into a 2ml collection tube.
  3. Apply the mixture to the QIAquick column to bind the DNA and centrifuge for 30 to 60 seconds at 13,000 rpm.
  4. Discard flow-through. Add 0.75 ml of Buffer PE to the spin column to wash and centrifuge for 30 to 60 seconds at 13,000 rpm.
  5. Discard flow-through and centrifuge for another 1 minute at 13,000 rpm.
  6. Place QIAquick spin column into a clean 1.5 ml microcentrifuge tube.
  7. Add 50 µl Buffer EB to the center of the membrane of spin column and centrifuge for 1 minute at 13,000 rpm.
  8. If analysing purified DNA using gel, add 1 volume of Loading Dye to 5 volumes of DNA.
  9. Store the PCR product either at 4°C or at -20°C.

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