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I have recently done some work analyzing the DNA content of nuclei
prepared from potato leaves, utilizing the protocol described by
Arumuganathan & Earle (1991). Briefly, the leaf tissue is sliced in a
magnesium sulfate buffer with dithiothreitol, Triton X-100, and
propidium iodide, filtered and centrifuged. The nuclei are then
stained with the above solution containing RNAase and CRBC as an
internal DNA standard prior to analysis on a FACScan.
I am now interested in trying to substitute another DNA stain, 7-AAD,
for the propidium iodide. However, my initial attempts have been
unsuccessful, even though I've tried it with and without the
dithiothreitol. Any other ideas or suggestions would be much
appreciated (Molecular Probes didn't have any).
My e-mail address is: cjgros@ccmail.monsanto.com;
Thanks for your time and consideration,
Cindy Gross
Monsanto EHL; 314/694-7948
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web