High-resolution mapping of satellite DNA using biotin-labeled DNA probes
| Autor: | P. R. Langer-Safer, Laura Manuelidis, David C. Ward |
|---|---|
| Rok vydání: | 1982 |
| Předmět: |
DNA nanoball sequencing
Satellite DNA Centromere Biotin In situ hybridization DNA Satellite Biology Chromosomes DNA sequencing Cell Line Immunoenzyme Techniques Mice chemistry.chemical_compound Nucleic acid thermodynamics Heterochromatin Animals Base Sequence Hybridization probe Nucleic Acid Hybridization Articles Glioma Cell Biology Molecular biology chemistry Biochemistry DNA |
| Zdroj: | The Journal of Cell Biology |
| ISSN: | 1540-8140 0021-9525 |
| DOI: | 10.1083/jcb.95.2.619 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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