sorting cells for RNA

Mike Salmon salmonm at rheuma.bham.ac.uk
Wed Apr 5 10:02:40 EST 1995


 
> 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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