Phenol extraction problem/phases not separating
Pete
pkursula at cc.oulu.fi
Mon Dec 2 03:19:10 EST 1996
On Fri, 29 Nov 1996, MARLENE SNYDER wrote:
> I am extracting DNA from frog toes collected in lysis buffer
> :SDS/Tris/EDTA/Sucrose - nothing out of the ordinary.
>
> I am using a standard phenol-chloroform extraction.
>
> Most of my extractions work as expected but about 1 in 8 refuses to separate;
> the lysis buffer and phenol mix. The buffer is the same in each sample and I
> don't understand why this is happening. So if anyone knows why and
> (more importantly) what I can do to make the phases separate so I can continue
> the extraction, I would appreciate hearing from you.
>
> Ian Paterson
>
In my experience, this may happen when your DNA concentration is very
high and thus your DNA solution is rather viscous. I would dissolve the
DNA in a larger volume and make the extraction then.
Pete
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