PEG purification of PCR products

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Overview

This is a simple and cost-effective protocol for purifying PCR products from primers and unused dNTPs. Purified products can be used in any downstream application, such as DNA sequencing.

To make PEG

  • 3g PEG 8000
  • 2.19g NaCl
  • Bring up to 15ml with good water.
  • Lasts for at least 30 days at 4C

Protocol

  • Add 1:1 volume of PEG to PCR product, for example into the reaction tube.
  • Vortex briefly.
  • Warm at 37degrees for 15minutes.
  • Microcentrifuge for 15minutes at max rpm.
  • Pipette off supernatant.
  • Wash pellet with exess 150ul 80% cold ethanol.
  • Spin for 10minutes at at max rpm.
  • Pipette off ethanol.
  • [Optional] Wash with 95% ethanol.
  • Dry down.
  • Re-suspend in 10 to 25ul of good water. Generally, I re-suspend 8μL of a PCR product in 12μL water.