Team:Newcastle/PCR purification

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(New page: =PCR Purification= ==Materials== * 1.5ml microcentrifuge tubes * 2ml collection tube * QIAquick columns * Buffer PB * Buffer EB * Buffer PE * DNA mixture from PCR ==Protocol== # Add 5 ...)
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Revision as of 10:10, 26 August 2010

PCR Purification

Materials

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

Protocol

  1. Add 5 volumes of 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.
  4. Discard flow-through. Add 0.75 ml of Buffer PE to the spin column to wash and centrifuge for 30 to 60 seconds.
  5. Discard flow-through and centrifuge for another 1 minute.
  6. Place QIAquick spin column into a clean 1.5ml microcentrifuge tube.
  7. Add 50 µl Buffer EB to the center of the membrane of spin column and centrifuge for 1 minute.
  8. If analysing purified DNA using gel, add 1 volume of Loading Dye to 5 volumes of DNA.