phenol extraction

Michael Witty mw132 at mole.bio.cam.ac.uk
Fri Mar 8 12:42:11 EST 2002


On 8 Mar 2002, marta varela rey wrote:

> I extracted DNA with phenol-water instead of phenol-tris by mistake.
> Should I go on with the procedure? Will I recover DNA from this
> extraction? In case I don=B4t, is there any way to recover the DNA?
> I would appreciate any help as soon as possible.
> Thanks,
> Marta.

Dear Marta,
          I have a feeling that DNA will move to the organic/phenol phase
if you have not used low pH buffered phenol.  Here is what I would do:

If you have retained the organic phase, add conc tris (or maybe tris
powder) to raise the pH to about 8 (measuring pH by taking drops from the
mixture and spotting onto pH paper, or you may damage the pH meter).  Then
take the new aqueous phase and continue.  (maybe continue with the acidic
aqueous phase too, just to be sure).

Good luck.  Mike.




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