Corum:PCR Purification

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  • SC 15:45, 19 June 2012 (EDT):


PCR Purification using Invitrogen's PureLink PCR Purification Kit.


  • DNA sample of volume V
  • PureLink PCR purification column
  • Binding buffer
  • Wash buffer
  • Sterile ddH2O


  1. Add 4V binding buffer (700 μL for concentrated DNA). Vortex and spin.
  2. Apply to column. Spin in tabletop centrifuge 10,000 rpm 1 min. Discard flowthrough.
  3. Apply 650 μL wash buffer to column. Spin 10,000 rpm 1 min. Discard flowthrough.
  4. Spin max speed 2 min to dry.
  5. Apply 100 μL sterile ddH2O to column membrane. Take care to not touch membrane with pipet tip.
  6. Incubate RT 10 min.
  7. Spin max speed 2 min to elute.
  8. Optional: Combine like samples and/or vacuum centrifuge at 60 °C to reduce volume. IMPORTANT: Time depends on initial and desired final volumes; samples should be checked frequently to prevent total evaporation.


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Relevant papers, books, and websites

  1. Invitrogen Life Technologies


or instead, discuss this protocol.