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A new method of in situ hybridization

Manning, Jerry E. and Hershey, N. Davis and Broker, Thomas R. and Pellegrini, Maria and Mitchell, Herschel K. and Davidson, Norman (1975) A new method of in situ hybridization. Chromosoma, 53 (2). pp. 107-117. ISSN 0009-5915.

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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.

Item Type:Article
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Additional Information:© 1975 Springer. Received 20 June 1975; Accepted 15 July 1975. Contribution number 5121 from the Department of Chemistry.
Subject Keywords:Polymer; Electron Microscope; Scanning Electron Microscope; Nucleotide; Gene Mapping
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Other Numbering System NameOther Numbering System ID
Caltech Division of Chemistry and Chemical Engineering5121
Issue or Number:2
Record Number:CaltechAUTHORS:20201020-103534448
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Official Citation:Manning, J.E., Hershey, N.D., Broker, T.R. et al. A new method of in situ hybridization. Chromosoma 53, 107–117 (1975).
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:106168
Deposited By: Tony Diaz
Deposited On:20 Oct 2020 20:47
Last Modified:20 Oct 2020 20:47

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