SDS extraction of whole cells

Michael Witty mw132 at mole.bio.cam.ac.uk
Fri Jul 27 15:57:46 EST 2001


On Fri, 27 Jul 2001, Warren Gallin wrote:

> Hi folks,
> 	I am trying to do a Western blot on a set of cell lines and am trying
> to keep the protein concentration as high as possible.  The problem is
> that the DNA released on boiling in SDS makes the sample impossible to
> pipette accurately.
> 	Aside from dilution, does anyone have a way to get the viscosity of
> this kind of extract down to a workable level?
>
> Thanks.
>
> Warren Gallin

Dear Warren,
           I had the same problem myself today.  My solution was to suck
the sample (250microlitre volume) through a narrow needle five or 10
times.  Be careful not to make foam.  If you do have a sonicator maybe
Louis Hom's advise is best though.  Mike





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