problems with phenol-choroform-isoamylalcohol extraction - (Aug/29/2008 )
Hi everyone! I was doing a Phenol chloroform extraction as usual, but this time didn´t work!!! 
. I didnt get any phase separation!!! could someone help me? I tried with buffer alone and the phenol mixed and it work fine, but not with my samples. Thanks!
-clutz-
I am assuming that the phenol-chloroform in question has a ratio of 25:24:1 = phenol:chloroform:isoamyl alcohol.
Hmmm... was your sample heavily contaminated with isopropanol? Isopropanol can disolve chloroform.
As for you sample, ethanol percipitate it, and you should be able to recover you DNA sample.
-perneseblue-
QUOTE (perneseblue @ Aug 29 2008, 09:02 AM)
I am assuming that the phenol-chloroform in question has a ratio of 25:24:1 = phenol:chloroform:isoamyl alcohol.
Hmmm... was your sample heavily contaminated with isopropanol? Isopropanol can disolve chloroform.
As for you sample, ethanol percipitate it, and you should be able to recover you DNA sample.
Hmmm... was your sample heavily contaminated with isopropanol? Isopropanol can disolve chloroform.
As for you sample, ethanol percipitate it, and you should be able to recover you DNA sample.
Thanks for your answer. no my sample didn´t contain any isopropanol, but I appreciate your suggestion of precipitate the whole thing. I´ll try that
-clutz-