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?