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Immunological method for mapping genes on Drosophila polytene chromosomes.
Open accessAbstract
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.
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Cited by
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Metadata
- Authors
- Langer-Safer, P. R., Levine, M., & Ward, D. C.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1982-07
- Volume
- 79
- Issue
- 14
- Pages
- 4381-4385
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2009993853
- Cited by
- 798
- References
- 0
- FWCI
- 32.62
- Type
- journal article
- Language
- en
- Open access
- green
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- Langer-Safer, P. R., Levine, M., & Ward, D. C. (1982). Immunological method for mapping genes on Drosophila polytene chromosomes. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.79.14.4381
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