Mapping small DNA sequences by fluorescence in situ hybridization directly on banded metaphase chromosomes

Journal Article · Wed Aug 01 00:00:00 EDT 1990 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.87.16.6223· OSTI ID:5725928

Fan, Y S; Davis, L M; Shows, T B ^[1]

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1. New York State Department of Health, Buffalo (USA)

A procedure for mapping small DNA probes directly on banded human chromosomes by fluorescence in situ hybridization has been developed. This procedure allows for the simultaneous visualization of banded chromosomes and hybridization signal without overlaying two separate photographic images. This method is simple and rapid, requires only a typical fluorescence microscope, has proven successful with DNA probes as small as 1 kilobase, is applicable for larger probes, and will greatly facilitate mapping the vast number of probes being generated to study genetic disease and define the human genome. Human metaphase chromosomes were prepared from phytohemagglutinin-stimulated lymphocyte cultures synchronized with bromodeoxyuridine and thymidine. Probes were labeled with biotin-dUTP, and the hybridization signal was amplified by immunofluorescence. Chromosomes were stained with both propidium iodide and 4{prime},6-diamidino-2-phenylindole (DAPI), producing R- and Q-banding patterns, respectively, allowing unambiguous chromosome and band identification while simultaneously visualizing the hybridization signal. Thirteen unique DNA segments have been localized to the long arm of chromosome 11 by using this technique, and localization of 10 additional probes by using radioactive in situ hybridization provides a comparison between the two procedures. These DNA segments have been mapped to all long-arm bands on chromosome 11 and in regions associated with neoplasias and inherited disorders.

• OSTI ID: 5725928
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Vol. 87:16; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
HETEROCHROMOSOMES
GENETIC MAPPING
BIOLOGICAL MARKERS
BUDR
DNA HYBRIDIZATION
FLUORESCEIN
FLUORESCENCE
HEREDITARY DISEASES
MAN
PHYTOHEMAGGLUTININ
SYNCHRONOUS CULTURES
AGGLUTININS
ANIMALS
ANTIBODIES
ANTIMETABOLITES
AROMATICS
AZINES
BROMOURACILS
CARBOHYDRATES
CARBOXYLIC ACIDS
CELL CULTURES
CHROMOSOMES
DISEASES
DRUGS
DYES
HEMAGGLUTININS
HETEROCYCLIC COMPOUNDS
HYBRIDIZATION
HYDROXY ACIDS
HYDROXY COMPOUNDS
LUMINESCENCE
MAMMALS
MAPPING
MITOGENS
MUCOPROTEINS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC BROMINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHENOLS
POLYPHENOLS
POLYSACCHARIDES
PRIMATES
PROTEINS
PYRIMIDINES
RIBOSIDES
SACCHARIDES
URACILS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques