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PMID: 6812046 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Immunological method for mapping genes on Drosophila polytene chromosomes.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 79 ·No. 14 ·1982-07-00 ·Pages 4381-5

Langer-Safer PR, Levine M, Ward DC

Abstract

A method is described for localizing DNA sequences hybridized in situ to Drosophila polytene chromosomes. This procedure utilizes a biotin-labeled analog of TTP that can be incorporated enzymatically into DNA probes by nick-translation. After hybridization in situ, the biotin molecules in the probe serve as antigens which bind affinity-purified rabbit antibiotin antibodies. The site of hybridization is then detected either fluorimetrically, by using fluorescein-labeled goat anti-rabbit IgG, or cytochemically, by using an anti-rabbit IgG antibody conjugated to horseradish peroxidase. When combined with Giemsa staining, the immunoperoxidase detection method provides a permanent record that is suitable for detailed cytogenetic analysis. This immunological approach offers four advantages over conventional autoradiographic procedures for detecting in situ hybrids: (i) the time required to determine the site of hybridization is decreased markedly, (ii) biotin-labeled probes are chemically stable and give reproducible results for many months; (iii) biotin-labeled probes appear to produce less background noise than do radiolabeled probes; and (iv) the resolving power is equal to and often greater than that achieved autoradiographically.

MeSH Terms
Animals Biotin/immunology Chromosome Mapping Chromosomes/ultrastructure Drosophila melanogaster/genetics Immunoenzyme Techniques Methods Nucleic Acid Hybridization
Chemicals
Biotin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Langer-Safer P R
Levine M
Ward D C
References (34)
34 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.79.14.4381
Published
1982-07-00
Pages
4381-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC346675
Subset
IM
Grants
NCI NIH HHS · CA-16038 · United States
NIGMS NIH HHS · GM-20124 · United States
External Links
PubMed source DOI: 10.1073/pnas.79.14.4381 PMC Full Text View journal details
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