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PMID: 6947272 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Transmission electron microscopic method for gene mapping on polytene chromosomes by in situ hybridization.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 78 ·No. 11 ·1981-11-00 ·Pages 7059-63

Wu M, Davidson N

Abstract

A transmission electron microscope method for gene mapping by in situ hybridization to Drosophila polytene chromosomes has been developed. As electron-opaque labels, we use colloidal gold spheres having a diameter of 25 nm. The spheres are coated with a layer of protein to which Escherichia coli single-stranded DNA is photochemically crosslinked. Poly(dT) tails are added to the 3' OH ends of these DNA strands, and poly(dA) tails are added to the 3' OH ends of a fragmented cloned Drosophila DNA. These probe--dA strands are hybridized in situ to polytene chromosome squashes. Gold spheres are linked to the hybridized probe--dA strands by A.T base pairing. The sphere positions relative to the chromosome bands can be observed by transmission electron microscopy. The method shows low background and high resolution.

MeSH Terms

Animals Chromosomes/ultrastructure DNA/genetics DNA Nucleotidyltransferases Deoxyadenine Nucleotides Drosophila Gold Microscopy, Electron Nucleic Acid Hybridization Poly A Poly T

Chemicals

Deoxyadenine Nucleotides Poly A Poly T poly(dA) Gold DNA DNA Nucleotidyltransferases

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Wu M

Davidson N

References (11)

11 references, click to expand

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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.78.11.7059

Published

1981-11-00

Pages

7059-63

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC349194

Subset

Grants

NIGMS NIH HHS · GM 20927 · United States

External Links

PubMed source DOI: 10.1073/pnas.78.11.7059 PMC Full Text View journal details

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Transmission electron microscopic method for gene mapping on polytene chromosomes by in situ hybridization.

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