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