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Chromosome-specific DNA Repeat Probes [electronic resource].

Published:
Washington, D.C. : United States. Dept. of Energy, 2006.
Oak Ridge, Tenn. : Distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy.
Additional Creators:
United States. Department of Energy, National Institutes of Health (U.S.), and United States. Department of Energy. Office of Scientific and Technical Information
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Summary:
In research as well as in clinical applications, fluorescence in situ hybridization (FISH) has gained increasing popularity as a highly sensitive technique to study cytogenetic changes. Today, hundreds of commercially available DNA probes serve the basic needs of the biomedical research community. Widespread applications, however, are often limited by the lack of appropriately labeled, specific nucleic acid probes. We describe two approaches for an expeditious preparation of chromosome-specific DNAs and the subsequent probe labeling with reporter molecules of choice. The described techniques allow the preparation of highly specific DNA repeat probes suitable for enumeration of chromosomes in interphase cell nuclei or tissue sections. In addition, there is no need for chromosome enrichment by flow cytometry and sorting or molecular cloning. Our PCR-based method uses either bacterial artificial chromosomes or human genomic DNA as templates with α-satellite-specific primers. Here we demonstrate the production of fluorochrome-labeled DNA repeat probes specific for human chromosomes 17 and 18 in just a few days without the need for highly specialized equipment and without the limitation to only a few fluorochrome labels.
Report Numbers:
E 1.99:lbnl--59871
lbnl--59871
Other Subject(s):
Note:
Published through SciTech Connect.
03/16/2006.
"lbnl--59871"
": 600301010"
Journal of Histochemistry and Cytochemistry 54 12 ISSN 0022-1554; JHCYAS FT
Weier, Heinz-Ulrich G.; Weier, Jingly Fung; Baumgartner, Adolf.
Ernest Orlando Lawrence Berkeley NationalLaboratory, Berkeley, CA (US)
Funding Information:
DE-AC02-05CH11231
NIHNIH: CA80792 ANDCA88258
L0083
View MARC record | catkey: 13809850