DNA, RNA, protein acetone precipitation? - (Dec/12/2007 )
Hi,
I sonificated (6x10sec on ice) total cells lysate (cells scraped in PBS with protease inhibitors). Then I added acetone (80% final volume, -20dgrC stored) to precipitate proteins. After centrifugation (21 000 x g) I got the acetone precipitate in a pellet.
1. My question is, if DNA and RNA are disrupted (in small fragments) by sonication are they in that pellet (after 21 000 x g)? And/or are they precipitated with acetone?
2. Other question, if I have proteins dissolved in mixture of phenol/ethanol, can I them effectively precipitate by acetone?
An evaporatio was not possible.
Thanks for suggestion,
vic
To make clear my question (1.):
I want to separate (RNA,) DNA from proteins - I want to have relatively pure protein pellet after acetone precipitation with small or no (RNA,) DNA contamination. I know that there are many possibilities how to separate DNA, RNA from proteins, but I would like to know whether what I have described should work – disruption of DNA, RNA by sonication and then acetone protein precipitation.
v.
I want to separate (RNA,) DNA from proteins - I want to have relatively pure protein pellet after acetone precipitation with small or no (RNA,) DNA contamination. I know that there are many possibilities how to separate DNA, RNA from proteins, but I would like to know whether what I have described should work – disruption of DNA, RNA by sonication and then acetone protein precipitation.
v.
I think that acetone selectively precipitates proteins.