TdT and PI in plants
Iona O'Brien (iobrien@marccri.marc.cri.nz)
Fri, 8 Dec 1995 13:54:25 GMT+1200
Help.
I am currently mapping chromatin condensation followed by nDNA
fragmentation in plant cells which have been treated with a range of
cytotoxic agents.
I am using the Boehringer Mannheim Cell Death detection kit which
is working well. However I want to double stain with propidium
iodide to follow the chromatin condensation and apoptotic peak versus
the TdT. When I try this using PBS I end up with peak cv's of 15-25%.
In comparison I obtain peak cv's of 2-5% when I replace the PBS with a
modified version of David Galbraith's buffer (45mM MgCl2,
45mM sodium citrate, 20mM MOPS). However using this buffer the
TdT gives poor results.
Does anyone have any suggestions on how to get around this problem or
will I have to continue to analyse separately.
I would also like to thank those people who offered advice on my
earlier question on how to measure calcium in plants. It worked
beautifully in the end. A special thanks to Geoff Osborne and Sabine
at ANU for all their help while I was working in their lab.
Thanks in advance,
Iona O'Brien
Scientist
Hort Research
Auckland
New Zealand.
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