SDS extraction of whole cells
cliodhnascanlon at hotmail.com
Sun Aug 5 19:10:29 EST 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?
Warren if you still wish to use the SDS method of extraction, why not add some DNase to your mixture...this always works for us. The high viscosity you mention is due to release of DNA on cell lysis. We always use a DNase premix called Benzonase from Merck...great stuff! However, we've also used DNaseI in a glycerol buffer. Just make sure to add the enzyme along with the SDS and you should have no problems! Good luck.
-------------- next part --------------
An HTML attachment was scrubbed...
More information about the Methods