Ploidy Analysis

Tim Kute tkute at wfubmc.edu
Fri Feb 6 14:25:10 EST 2004


Have you tried mixing the control cells with you triploid cells. This
will hopefully correct for changes in PI staining since both the control
cells and the triploid are exposed to the same PI concentration.
Tim

-----Original Message-----
From: Cogswell, Andrew [mailto:CogswellA at mar.dfo-mpo.gc.ca] 
Sent: Thursday, February 05, 2004 7:43 AM
To: cyto-inbox
Subject: Ploidy Analysis


Dear Flowers,

I am currently assessing triploidy induction rates using Propidium
iodide as a DNA specific flourecsent dye.  I find that when I run
controls containing diploid nuclei, the peak on my histograms tends to
shift significantly to the right or left of where I set it on the FL2
(x) axis from one run to another.  Therefore, when I run triploid
samples I cannot be sure (without some kind of internal standard) where
the actual peak lies, and whether it is in fact diploid.  Any ideas for
what I might do to standardize samples so that there is no question of
my samples ploidy level? 

I am a new user, so please feel free to elaborate.

Andrew T. Cogswell
Science Branch, Maritimes Region
Invertebrate Fisheries Division
Fisheries and Oceans Canada
1 Challenger Drive
Halifax, NS
B2Y 4A2
Phone: 902-426-4353 




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