Hallo Louis, I think this paper might help you,
GENES & DEVELOPMENT 14: 212-223
Dynamic organization of chromosomal DNA in Escherichia coli.
Hironori Niki, Yoshiharu Yamaichi, Sota Hiraga.
(You can get it online if you subscribe www.genesdev.org)
E-mail: niki@gpo.kumamoto-u.ac.jp
I allways measure my bacteria with a jet in air FACSVantage, and I make my
DNA analysis by staining with Hoechst 33258 or DAPI, that allows me to
differenciate the alteration in the number of chromosomes between
exponential (multiple) and stationary (1N and 2N) phases in a batch culture
of E. coli.
I hope this information is useful for you.
Good luck!
-----------------------------------------
Elena Soriano
c/o Institut für Angewandte Mikrobiologie
Univ.f.Bodenkultur Wien
Tel: +43 1 36006 6241
Fax: +43 1 36 97 615
http://www.boku.ac.at/iam
------------------------------------------
To: cyto-inbox
I have a request from a person for flow cytometric help in identifying the
number of chromosomes replicating DNA in a bacterium. He has altered
proteins he/they think are involved in bacterial chromosome replication so
they want to compare the parent strain to the altered strain. He wants to
be able to establish significantly altered chromsomal replication patters
having 3 or more replication forks.
Any body out there in the US doing this? How about world wide?
Has anybody out there tried to do this on a jet in air FACS unit? (You may
laugh now. If by some miracle somebody did get even close to making this
work--drop me a line will you?)
Does anybody have a microscope based FACS unit that will do this? Partec
or other variety?
Any useful comment from experience will be greatly appreciated.
Louis King
Louis King, Ph.D.
Michigan State University Flow Cytometry Center
Department of Biochemistry and Molecular Biology, Rm 419
Michigan State University
East Lansing, MI 48824
Phone: 517-355-1536
FAX: 517-353-9334
E-Mail: kingL@pilot.msu.edu
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