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phenol in alcohol precip

phenol in alcohol precip - Protocols and Methods Forum

phenol in alcohol precip - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 04-21-2009, 01:38 PM
Ed Siefker
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Default phenol in alcohol precip



Hi. I was a little hasty in doing some phenol/chloroform
preps of genomic DNA and apparently didn't remove all
the phenol before adding ethanol to precipitate. How can
I recover from this?

I considered extracting with chloroform again, but I don't
know if the phases will still separate with ethanol in there.
Also, the DNA isn't in solution, so I'm guessing it would
collect at the interface. If there were more DNA there,
I'd just fish it out with a pipette tip, but it's a small prep.

Any thoughts? Thanks a bunch
-Ed

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Old 04-21-2009, 09:09 PM
WS
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Default phenol in alcohol precip

Dear Ed,

don't worry. Just rinse your pellet a few times with EtOH (and spin
down shortly to make sure you don't loose the pellet).. If you plan
downstream processing with enzymes, make sure all phenol has been
removed. You might dare to check thesmell; your nose should turn out
to be a very sensitive GC system with a neuronal detector attached to.
Chloroform might not be that good choice, even that DNA doesn't
dissolve in it, (regardless if there is EtOH and or phenol present),
as it does not dissolve phenol as well as ethanol does. You might
remember: the DNA (and other water soluble stuff) is in the phenol
phase, lipids are in the chloroform phase and proteins are inbetween.

Have fun!

Wo
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