Chromosome-specific DNA Repeat Probes

Baumgartner, Adolf; Weier, Jingly Fung; Weier, Heinz-Ulrich G.

doi:10.1369/jhc.6A6974.2006

Chromosome-specific DNA Repeat Probes

Journal Article · Thu Mar 16 04:00:00 EST 2006 · Journal of Histochemistry and Cytochemistry

DOI:https://doi.org/10.1369/jhc.6A6974.2006· OSTI ID:919765

Baumgartner, Adolf; Weier, Jingly Fung; Weier, Heinz-Ulrich G.

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 {alpha}-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.

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Research Organization:: Ernest Orlando Lawrence Berkeley NationalLaboratory, Berkeley, CA (US)

Sponsoring Organization:: USDOE Director, Office of Science; National Institutes ofHealth

DOE Contract Number:: AC02-05CH11231

OSTI ID:: 919765

Report Number(s):: LBNL--59871; BnR: 600301010

Journal Information:: Journal of Histochemistry and Cytochemistry, Journal Name: Journal of Histochemistry and Cytochemistry Journal Issue: 12 Vol. 54; ISSN JHCYAS; ISSN 0022-1554

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
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HUMAN CHROMOSOMES
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Chromosome-specific DNA Repeat Probes

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