cell cycle in bacteria using PI

From: Luciana Andrade <lnsa@usp.br>
Date: Fri Jul 04 2008 - 16:32:37 EDT
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,
Luciana
Received on Fri Jul 4 18:58:00 2008

This archive was generated by hypermail 2.1.8 : Wed Jan 31 2007 - 03:12:00 EST