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-