Re: cellular contents separation protocol
Gautam Sarath (gsarath@unlinfo.unl.edu)
Tue, 3 Mar 1998 18:03:29 -0600 (CST)
>Dear all on ABRF:
>
>Can anyone give me some leads for protocols to isolate SIMULTANEOUSLY DNA,
>RNA and protein from cells...probably yeast , Pls?
>
>I would like to use this in an application where I am "feeding" isotope
>oxygen (heavy water) to yeast, then harvest the DNA, RNA and Protein for
>those contents that successfully incorporated the isotope ....
>
>Any clues will be appreciated.
>
>Sincerely,
>
>Winnell Newman
>
>Winnell H. Newman Director, Nuclei Acids Facility
>919-515-3463 office North Carolina State University
>919-515-3355 fax Dept of Genetics Box 7614 (mail)
>winnell@ncsu.edu 3513 Gardner Hall (shipping)
> Raleigh, NC 27695
Winnell:
You might try the regular Phenol/chloroform extraction used for preparing
DNA. This should precipitate all of the proteins, which can then be washed
with cold acetone and freeze-dried. You will then obtain DNA + RNA in the
aqueous phase. My expertise is with proteins, but I am sure that there are
classical (non-RNAse) means of obtaining enriched fractions for RNA and
DNA, either based on size (sedimentation) or on columns, which will bind
the double stranded DNA, much tighter than the single stranded RNA. Gautam
Sarath
Gautam Sarath
N-226, Beadle center
Protein Core Facility
Department of Biochemistry
University of Nebraska-Lincoln
Lincoln, NE 68588-0664
Phone: 402-472-2928
FAX: 402-472-7842