Corum:PCR Purification

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  • SC 15:45, 19 June 2012 (EDT):

Contents

Overview

PCR Purification using Invitrogen's PureLink PCR Purification Kit.

Materials

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

Procedure

  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.

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

  1. List troubleshooting tips here.
  2. You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
  3. Anecdotal observations that might be of use to others can also be posted here.

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References

Relevant papers, books, and websites

  1. Invitrogen Life Technologies

Contact

or instead, discuss this protocol.

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