Phenol extraction/proteins/western

Frank O. Fackelmayer fof1 at chclu.chemie.uni-konstanz.de
Wed Sep 9 05:07:17 EST 1998


Yes.
You can recover the interphase together with the organic phase (everything
usually left behind in DNA preps), and add methanol to precipitate the
proteins. 4 volumes (i.e. 400ul methanol for 100ul organic phase) of methanol
is ok. Vortex vigorously, then spin down proteins for 5min in a desktop
centrifuge, max. speed. There will be a precipitate and a liquid phase. No
phase separation should be evident. If you DO see a phase separation, add more
methanol until phases do not separate any more. Remove liquid supernatant, air
dry pellet, then dissolve it in SDS sample buffer. In case the sample does not
dissolve easily, sonicate it briefly before boiling. Spin the sample for 5min
at max. speed to remove undissolved material that could cause streaking in the gel.

Hope this helps,
Frank

Nemo Peeters wrote:
> 
> Hi,
> Can i use proteins that are at the interface of a phenol/chlorophorm
> nucleic acid extraction to do some western blots ?. I have no idea how to
> recover them, did some of you try this kind of experiment ?
> 
> regards



More information about the Methods mailing list