no waterphase in phenolextraction

LoPony ctfrench at wwisp.com
Tue May 30 23:27:11 EST 2000


Make sure the phenol is saturated with buffer-

Try adding 1/4 to 1 vol of chloroform to the prep after mixing the phenol
and then centrifuge.  I've had this experience myself in some RNA
extractions.  A little chloroform won't hurt anything and adds density of
course.

Good luck.

--
==============================
Christopher Todd French
UAB Dept. of Comparative Medicine
1670 Univ. Blvd, VH 422
1530 3rd Ave. S
Birmingham, AL 35294-0019

==============================

"Gerd " <gerdn at ibg.uit.no> wrote in message
news:8gg281$1o1$1 at news.uit.no...
> Hei,
> When intending to isolate genomic bacterial DNA by a method involving
> lysozyme -> SDS -> phenolextraction, a person in our lab experienced a
> strange phenomenon. Everything appeared normal until the phenol step -
> after adding the phenol, it sank to the bottom, ok, then he mixed the
> phases, sentrifuged - and found no waterphase.
> Adding NaCl helped produce some waterphase, but after finnishing the
> procedure it was clear that no DNA were recovered.
> What could be the explanation?
> He has used the same precedure rutinely (although a while ago),
> phenol was well buffered, solutions were personal (no mixup likely)
>
> Any suggestions?







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