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A new method of in situ hybridization
- Source :
- Chromosoma. 53:107-117
- Publication Year :
- 1975
- Publisher :
- Springer Science and Business Media LLC, 1975.
-
Abstract
- A new method for gene mapping at the chromosome level using in situ hybridization and scanning electron microscopy is described and has been applied to mapping the rRNA genes of Drosophila melanogaster. Biotin is covalently attached to Drosophila rRNA via a cytochrome c bridge at a ratio of one cytochrome-biotin per 130 nucleotides by a chemical procedure. Polymethacrylate spheres with a diameter of ca. 60 nm are prepared by emulsion polymerization and are covalently attached to the protein avidin at a ratio of 5-20 avidins per sphere. The biotin-labeled rRNA is hybridized to denatured DNA in a chromosome squash. Upon incubation with a sphere solution, some of the biotin sites become labeled with spheres because of the strong non-covalent interaction between biotin and avidin. The chromosome squash is examined in the scanning electron microscope (SEM). Polymer spheres, which are visible in the SEM, are observed to label the nucleolus, where the rRNA genes are located.
- Subjects :
- Nucleolus
Biotin
Cytochrome c Group
Nucleic Acid Denaturation
law.invention
chemistry.chemical_compound
Gene mapping
law
Methods
Genetics
Animals
Methylmethacrylates
Nucleotide
Genetics (clinical)
chemistry.chemical_classification
biology
Chromosome Mapping
Nucleic Acid Hybridization
DNA
Ribosomal RNA
Avidin
Microspheres
Drosophila melanogaster
Genes
Biochemistry
chemistry
RNA, Ribosomal
Microscopy, Electron, Scanning
biology.protein
Biophysics
Electron microscope
Subjects
Details
- ISSN :
- 14320886 and 00095915
- Volume :
- 53
- Database :
- OpenAIRE
- Journal :
- Chromosoma
- Accession number :
- edsair.doi.dedup.....4feca918db108b4b7fb1f9f040c526e1