I need to purify PCR product to further proceed for cloning, I dont have PCR purification kit so have to do manually. if I do Phenol:Chloroform:Isoamyl Alcohol 25:24:1 purification and ethanol precipitation, will it remove Polymerase also ????
Dear Priti,
You can use phenol chloroform extraction. But for many tasks the extraction from the gel could be better to remove matrix DNA. For that it is possible to centrifuge through the cotton clamp the gel fragment with PCR product after electrophoresis if no kits are available.
Phenol-chloroform extraction will certainly denature polymerase and inactivate it. It will do so for nearly all proteins. However you need to be careful to ensure you actually recover your PCR product after doing so, and that you remove all traces of the phenol before moving on to the next step.
- Badges
- Science method