Rapid generation of chromosome-specific alphoid DNA probes using the polymerase chain reaction

Ian Dunham, Christoph Lengauer, Thomas Cremer, Terry Featherstone

Research output: Contribution to journalArticle

Abstract

Non-isotopic in situ hybridization of chromosome-specific alphoid DNA probes has become a potent tool in the study of numerical aberrations of specific human chromosomes at all stages of the cell cycle. In this paper, we describe approaches for the rapid generation of such probes using the polymerase chain reaction (PCR), and demonstrate their chromosome specificity by fluorescence in situ hybridization to normal human metaphase spreads and interphase nuclei. Oligonucleotide primers for conserved regions of the alpha satellite monomer were used to generate chromosome-specific DNA probes from somatic hybrid cells containing various human chromosomes, and from DNA libraries from sorted human chromosomes. Oligonucleotide primers for chromosome-specific regions of the alpha satellite monomer were used to generate specific DNA probes for the pericentromeric heterochromatin of human chromosomes 1, 6, 7, 17 and X directly from human genomic DNA.

Original languageEnglish (US)
Pages (from-to)457-462
Number of pages6
JournalHuman Genetics
Volume88
Issue number4
DOIs
StatePublished - Feb 1992
Externally publishedYes

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DNA Probes
Human Chromosomes
Chromosomes
Polymerase Chain Reaction
DNA Primers
Chromosomes, Human, Pair 6
Heterochromatin
Hybrid Cells
Chromosomes, Human, Pair 1
Interphase
Metaphase
Fluorescence In Situ Hybridization
Gene Library
In Situ Hybridization
Cell Cycle
DNA

ASJC Scopus subject areas

  • Genetics(clinical)
  • Genetics

Cite this

Rapid generation of chromosome-specific alphoid DNA probes using the polymerase chain reaction. / Dunham, Ian; Lengauer, Christoph; Cremer, Thomas; Featherstone, Terry.

In: Human Genetics, Vol. 88, No. 4, 02.1992, p. 457-462.

Research output: Contribution to journalArticle

Dunham, Ian ; Lengauer, Christoph ; Cremer, Thomas ; Featherstone, Terry. / Rapid generation of chromosome-specific alphoid DNA probes using the polymerase chain reaction. In: Human Genetics. 1992 ; Vol. 88, No. 4. pp. 457-462.
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