FACS sorting into what solution for RNA isolation? - (Jun/25/2008 )
I'm doing a FACS sort of a modest amount of cells (30,000-100,000), with the ultimate goal of isolating the RNA and using it for qRT-PCR. I'm undecided as to the best solution to sort the cells into. Trizol? RNAlater? Normal media that keeps cells happy? The first time I sort I'll have time to isolate the RNA directly after the sort [but even still, some of the populations are rare and take a while to get an adequate amount], though that may not be true at later dates. Also, I've attempted to get cells out of RNAlater before and have had trouble- even with the 1:1 dilution with PBS.
Any suggestions would be greatly appreciated!
I sort my cells into FACS buffer and try and kepe them happiest by keeping them on ice and doing the sort etc as quick as possible.
I sort the cells in FACS buffer, as soon as the sample is sorted I spin down the cells and resuspend them in the cell culture medium of choice.
Then I let them rest ON in the incubator, before I harvest and prep protein and RNA.