PEG purification of PCR products
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
- 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.