Hi all, I am trying with no success to get sharp DNA peaks using PI (propidium iodide) in bacteria (Caulobacter crescentus). I fixed them with ethanol (70%) and ressuspended in PI solution (0.1% Triton X-100, 20ug/mL PI and 200ug/mL RNase A) for 30-60 minutes at room temperature. I am using side scatter (log scale) versus red fluorescence (lin/log scales) to visualize the population. In the histograms (lin and log) I do not get distincts peaks (G1 and G2) even though when I acquire a synchronized cell culture at different cell cycle phases. I saw many papers that use Sytox from Invitrogen to stain DNA bacteria. Is it possible to get G1/G2 DNA peaks using PI in bacteria? How can I get a better DNA peak resolution in the histograms? I am using a flow cytometry from Guava technologies (Easy Cyte plus). Thanks in advance, LucianaReceived on Fri Jul 4 18:58:00 2008
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