a question on cell cycle analysis with PI

Scott Tighe stighe at zoo.uvm.edu
Tue Mar 26 16:46:20 EST 2002


Yalin:

0.1% triton X will certainly make the cells permeable but will not fix
them. If you do not include RNase, then RNA  will also be stained, which
will influence your DNA cell cycle data. If you choose to add RNase A,
use at a final concentration of 0.25 mg/ml in your samples. You can take
a look at our protocol if you wish; it is for general DNA cell cycle
staining of mammalian cells. Our protocol employs the use of ethanol
fix/perm, but certainly can be used with Triton X.
http://www.vermontcancer.org/Research/Cores/FlowProtocols.html

Sincerely

Scott Tighe
Vermont Cancer Center
Flow Cytometry Core Lab

Yalin Guo wrote:
>
> 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.
>
> Yalin



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