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Hi Deb,
I've used Brdu to reveal the nascent DNA in the thymine-deprived E.coli.
cells
Monosov, E.Z., Toukdarian A., Helinski D., and S.J. Singer.(1992)
Immunoelectron microscopy of deoxyribonucleic acid in procaryotic cells.
Acta. Histochem. Cytochem. Vol.25, No.1&2, pp 137-141.
Unfortunately, I don't have the PDF , but, if you are interested, I can
Fax you a copy or I can send you a preprint.
Best.
Ed
Deb Berglund wrote:
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi everybody,
>
> Since we are talking about bacteria in flow, I have a new question for
> you all. I have a user who wants to do BRDU uptake in bacteria. She
> has had success with the procedure in biofilms using confocal and an
> adapted-from-mammalian labelling prcedure, but when we rip apart the
> biofilms and try flow, we seem to be analyzing debris, not individual
> bacteria. I have been surprisingly quite successful in looking at
> bacteria in flow, but think that the BRDU protocol is causing enough
> cells to lyse/leak that we are seeing labelled DNA clumps. I tossed a
> DNA/RNA stain on the samples for threshold and think that we are still
> seeing non-bacteria debris of varying sizes.
>
> Has anyone out there done BRDU uptake in bacteria with flow, and would
> you send me your protocol?
>
> Thanks in advance.
>
> Deb Berglund
> Montana State University
>
>
--
Edward Monosov, Ph.D.
Director , Cell Imaging & Histology
The Burnham Institute
10901 N. Torrey Pines Rd,
La Jolla, CA 92037
Ph: (858)646-3100 ext. 3206
Fax: (858)646-3196
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