BrdU incorporation ?'s

A. Coonrod starchie at u.washington.edu
Thu Jul 18 14:14:25 EST 1996


I'm interested in using a bromo-DNA product as a probe to study
transfection mechanisms in cells genetically deficient in lysosomal
function or in the presence of lysosomal protease inhibitors.

A short technical report  recently publisheded in BIOTECHNIQUES (21:82-86
July 1996) <<Application of 5-Bromo-2'Deoxyuridine as a Label for In Situ
Hybridization in Chromosome Microdissection and . . . >> gives no
references in the abstract when mentioning, "An especially useful
application is that it [BrdU] may be incorporated into probe DNA while . .
.  plasmids in bacteria are growing in the presence of BrdU."

Does anyone have protocol references for incorporating BrdU into probe
plasmid DNA grown in bacteria,  or perhaps, a supply of E. coli thymine
auxotrophs lying around?

Also,  I purchased some BrdUTP from SIGMA (for PCR incorporation
of BrdU),  and would like to have an alternate, more reliable source.
Any more known suppliers of bromodeoxyuridines?
Thanks for your time.

Archie Coonrod <starchie at u.washington.edu>
M. Horwitz Lab
Div of Medical Genetics
University of Washington




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