Ammonium Acetate Protein Precipitation

BACKD at QUCDN.QueensU.CA BACKD at QUCDN.QueensU.CA
Sun Jun 28 10:30:14 EST 1992


We've used 2.5M ammonium acetate for protein ppt'ion for several years with
good success. We use it to: 1) remove bulk protein (and a lot of RNA) from
plasmid preps prior to CsCl, 2) clean up plasmid and lambda preps that don't
digest well with restricition enzymes (presumably the high salt also aids in
stripping proteins bound to the DNA).
Plasmid minipreps taken through an ammonium acetate + EtOH ppt'ion usually
can be digested with restriction endonucleases without phenol extraction.
However, I don't know if that step gets rid of RNase. Presumably your
proteinase-K step should get kill RNase. I think it worth trying with
one or two preps. Let me know how it turns out.

Don Back
Dept. Biochem.
Queen's University
Kingston, Ontario. CANADA



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