a question on cell cycle analysis with PI
d_coder at MSN.com
Sun Mar 31 11:09:34 EST 2002
This protocol was designed to be used with live cells, although I have used
it with a wide variety of animal and plant cells that have been fixed. With
live cell suspensions, nuclei are freed by hypotonic lysis. RNase does need
to be added as PI will label double stranded RNA as well as double stranded
DNA. In the abscence of RNase, peak resolution is less precise. (RNase is
added at 30 Kuenitz units/mL.) In the technique described by Tate et a1.
(see Cytometry 4:211-215, 1983 for detailed protocol), NaCl is added to gain
normal tonicity before analysis as Na increases PI fluorescence intensity.
David M. Coder, Ph.D.
Consultant in Cytometry
email: d_coder at msn.com or dcoder1 at hotmail.com
----- Original Message -----
From: "Yalin Guo" <guo at mm11.ukl.uni-freiburg.de>
Sent: Tuesday, March 26, 2002 6:12 AM
Subject: a question on cell cycle analysis with PI
> I am doing a cell cycle analysis using cultured and non-cultured
> human CD34+ cells. I got a protocol with PI staining (0.1% Na
> Citrate, 0.1% Triton X-100, 20 ug/ml PI), but without fixation
> and RNase. I should be very grateful to get any suggestions for
> this method.
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