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phenol/CH3 extraction--no emulsion?

Ed Siefker via methods%40net.bio.net (by ebs15242 from gmail.com)
Mon Feb 11 13:58:39 EST 2013


I am doing a phenol/CH3 extraction with Life Technologies Totally RNA kit.
I found that I had reached the bottom of the bottle of phenol that comes
with the kit, and used another bottle that came from Fisher.  I used the
Fisher phenol for cleaning up a maxi-prep last week, so I didn't think
there would be a problem.

However, when I added the Fisher phenol to the cell lysate, they
did not form an interface or emulsion.  I got thick, easily visible
schlieren patterns in the tube which disappeared after heavy
vortexing leaving a clear solution.  Something about the Totally
RNA denaturation solution makes it miscible with this phenol.


I've been doing this for 7 years now and I haven't seen anything
like this before.   I'd like to recover these samples if I can.  What's
more likely to be successful? Addition of water or addition of phenol
to break the interface?  I have plenty of acid phenol from the kit,
so I can do either.   Or maybe addition of 50/50 h2o/phenol?



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