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PMID: 6754749 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

High-resolution mapping of satellite DNA using biotin-labeled DNA probes.

The Journal of cell biology ·Vol. 95 ·No. 2 Pt 1 ·1982-11-00 ·Pages 619-25

Manuelidis L, Langer-Safer PR, Ward DC

Abstract

We have developed a novel method for high resolution mapping of specific DNA sequences after in situ hybridization. DNA probes, labeled with biotin-nucleotides in conventional nick-translation reactions, are hybridized to cytological preparations and detected with affinity-purified rabbit antibiotin antibodies followed by antibodies to rabbit IgG that are conjugated to fluorescent or enzymatic reagents. Using peroxidase labeled anti-rabbit IgG, we are able to detect and localize specific sequences at both the light and electron microscopic levels. Initial studies were done with repeated DNA sequences previously mapped by light microscope autoradiography to assess the fidelity and resolution of this method. An analysis using biotin-labeled mouse satellite DNA is presented here.

MeSH Terms
Animals Base Sequence Biotin/immunology Cell Line Centromere/analysis Chromosomes/analysis,ultrastructure DNA, Satellite Glioma Heterochromatin/analysis Immunoenzyme Techniques Mice Nucleic Acid Hybridization
Chemicals
DNA, Satellite Heterochromatin Biotin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Manuelidis L
Langer-Safer P R
Ward D C
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22 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.95.2.619
Published
1982-11-00
Pages
619-25
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2112973
Subset
IM
Grants
NCI NIH HHS · CA-15044 · United States
NCI NIH HHS · CA-16038 · United States
NIGMS NIH HHS · GM-20124 · United States
External Links
PubMed source DOI: 10.1083/jcb.95.2.619 PMC Full Text View journal details
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