Re: sorting cells for RNA

Mike Salmon (salmonm@rheuma.bham.ac.uk)
Wed, 5 Apr 1995 15:02:40 +0000


> I will soon be sorting cells to obtain RNA from sorted cell populations. I
> was wondering if anyone had dealt with such issues as rinsing the lines with
> RNAse inhibitors, sorting directly into solutions containing RNAse
> inhibitors, keeping the instrument cold, etc.

I must admit we just sort cells in a sterile fashion as we would for
culture experiments, provided the cells remain intact there is not a
big problem. There again if we extract RNA from cultured cells we
don't do RNAse-free cultures, and we certainly don't have any
RNAse-free people for fresh isolates. The main thing seems to be to
wash the cells well and make sure the RNAse protection in the lysis
buffer is adequate.

Have fun

Mike

...........................................................
Mike Salmon
Department of Rheumatology
The Medical School
University of Birmingham
Birmingham B15 2TT
United Kingdom
Tel: 44 (0)121 414 6780
Fax: 44 (0)121 414 6794


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