Hi, The question is about whether proteins precipitate when I precipitate DNA (approx 3500 bp) with PEG600. Anyone looked up this or have experience with it? I have a pure DNA that I cleaved and I want to remove the Restriction enzyme from it.
I don't want to use kits... as I have so much DNA that it is impractical. I can't use phenol chloroform due to new health and safety regulations at the department. So it appears I might end up with PEG precipitation. I have no problems with the procedure, but can't find any info if the proteins (i.e. Restriction enzymes or PCR polymerases) can be efficiently removed from the DNA during PEG precipitation?
Also, does an ethanol wash of the pellet suffecient to remove the salts associated with PEG precipitation?
Thank you for all your help.
Hi Wassiti
is precipitation with PEG the only applicable way for you to precipitate your DNA? if it is, I should state that PEG also could precipitates proteins and I think it is not a good choice for your case. however, it depends on PEG size and the nature of target protein. why do not you use other possible ways such as precipitation of enzyme with heating or ammonium sulfate treatment?
What do you intend to do with the DNA once purified? Most RE can be heat inactivated.
I agree; PEG will bring down proteins, which may remain active. What about using higher levels of EDTA? REs are metalloenzymes and will only cope with so much EDTA in the system.
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