Strand-specific fluorescence in situ hybridization: The CO-FISH family

S. M. Bailey, E. H. Goodwin, Michael Cornforth

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

The ability to prepare single-stranded chromosomal target DNA allows innovative uses of FISH technology for studies of chromosome organization. Standard FISH methodologies require functionally single-stranded DNAs in order to facilitate hybridization between the probe and the complementary chromosomal target sequence. This usually involves denaturation of double-stranded probes to induce temporary separation of the DNA strands. Strand-specific FISH (CO-FISH; Chromosome Orientation-FISH) involves selective removal of newly replicated strands from DNA of metaphase chromosomes which results in single-stranded target DNA. When single-stranded probes are then hybridized to such targets, the resulting strand-specific hybridization is capable of revealing a level of information previously unattainable at the cytogenetic level. Mammalian telomeric DNA consists of tandem repeats of the (TTAGGG) sequence, oriented 5′→3′ towards the termini of all vertebrate chromosomes. Based on this conserved structural organization, CO-FISH with a telomere probe reveals the absolute 5′→3′ orientation of DNA sequences with respect to the pter→qter direction of chromosomes. Development and various applications of CO-FISH will be discussed: detection of cryptic inversions, discrimination between telomeres produced by leading-versus lagging-strand synthesis, and replication timing of mammalian telomeres. Copyright c 2004 S. Karger AG, Basel.

Original languageEnglish (US)
Pages (from-to)14-17
Number of pages4
JournalCytogenetic and Genome Research
Volume107
Issue number1-2
DOIs
StatePublished - 2004

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Carbon Monoxide
Fluorescence In Situ Hybridization
Chromosomes
Telomere
Single-Stranded DNA
DNA
Tandem Repeat Sequences
Aptitude
DNA Probes
Metaphase
Cytogenetics
Vertebrates
Technology

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Strand-specific fluorescence in situ hybridization : The CO-FISH family. / Bailey, S. M.; Goodwin, E. H.; Cornforth, Michael.

In: Cytogenetic and Genome Research, Vol. 107, No. 1-2, 2004, p. 14-17.

Research output: Contribution to journalArticle

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