Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization

D. Pinkel, T. Straume, Joe Gray

Research output: Contribution to journalArticle

2725 Citations (Scopus)

Abstract

This report describes the use of fluorescence in situ hybridization for chromosome classification and detection of chromosome aberrations. Biotin-labeled DNA was hybridized to target chromosomes and subsequently rendered fluorescent by successive treatments with fluorescein-labeled avidin and biotinylated anti-avidin antibody. Human chromosomes in human-hamster hybrid cell lines were intensely and uniformly stained in metaphase spreads and interphase nuclei when human genomic DNA was used as a probe. Interspecies translocations were detected easily at metaphase. The human-specific fluorescence intensity from cell nuclei and chromosomes was proportional to the amount of target human DNA. Human Y chromosomes were fluorescently stained in metaphase and interphase nuclei by using a 0.8-kilobase DNA probe specific for the Y chromosome. Cells from males were 40 times brighter than those from females. Both Y chromosomal domains were visible in most interphase nuclei of XYY amniocytes. Human 28S ribosomal RNA genes on metaphase chromosomes were distinctly stained by using a 1.5-kilobase DNA probe.

Original languageEnglish (US)
Pages (from-to)2934-2938
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume83
Issue number9
StatePublished - 1986
Externally publishedYes

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Cytogenetic Analysis
Metaphase
Fluorescence
Interphase
Chromosomes
DNA Probes
DNA
28S Ribosomal RNA
Chromosomes, Human, Y
Avidin
Hybrid Cells
Y Chromosome
Human Chromosomes
Biotin
Cell Nucleus
Fluorescence In Situ Hybridization
rRNA Genes
Chromosome Aberrations
Cricetinae
Anti-Idiotypic Antibodies

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. / Pinkel, D.; Straume, T.; Gray, Joe.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 83, No. 9, 1986, p. 2934-2938.

Research output: Contribution to journalArticle

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