atutter at aim.salk.edu
Sat Jun 1 13:28:31 EST 1996
> PROUET wrote:
> We have now another problem. It seems that when you mix your DNA (PCR
> products of about 3 kb) with a new phenol (that is supposed to be
> equilibrated in T.E.) you loose the DNA !!! It worked just fine for years
> and now it looks like it could come from a new phenol batch. Did anybody
> experienced such problems ? Or what is your idea about that ? All
> suggestions are welcome ! (The first idea is that we will not use that
> phenol any longer !!!).
I always equilibrate phenol with 0.1M Tris, pH 7.9. Is it possible that you did not
equilibrate the phenol to a pH of >7.0? At acid pH, DNA will enter the phenol layer, and
not the aqueous layer. Luckliy, I did not learn this the hard way!
Let me know if it works!
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