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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