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Percentage of SDS to denature RNases? - (Oct/06/2005 )

Currently I've been having trouble with RNA degradation of hyphal samples versus yeast samples. I want to wash the cells in a solution containing SDS before freezing to get rid of any RNases and am wondering what concentration of SDS to use. Any suggestions?

-orca-

interesting question. SDS will lyse cells, is this a problem for you?

-John Buckels-

QUOTE (John Buckels @ Oct 7 2005, 01:13 AM)
interesting question. SDS will lyse cells, is this a problem for you?


I don't want the cells to lyse. I'm hoping the cell walls keep the cells from lysing, but I want to get rid of any proteins that stick to the outside, from the serum in the medium I am using to grow the cells. Any suggestions?

-orca-

SDS by its very nature will lyse cells

I would suggest trying washing these cells with a range of SDS solutions from 0.01% up to 5%, looking under the microscope to see if the cells are still intact then assessing whether these proteins are still stuck to the cell walls

-John Buckels-