Methods of fluorescence in situ hybridization on extended DNA fibers (Fiber-FISH)

Jason G. Walling, Robert M. Stupar, Jiming Jiang

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

Fiber-fluorescence in situ hybridization (fiber-FISH) is a cytogenetic technique that can be used to visualize DNA probes on extended DNA fibers. Fiber-FISH provides a significantly higher mapping resolution than do the chromosome- or chromatin-based cytogenetic mapping techniques. Here we report a simple procedure for performing fiber-FISH. In general, we consider the overall protocol to comprise four distinct steps, each of which is critical to the success of the experiment: the isolation of nuclei from plant tissue, the extension of fibers on glass slides, the hybridization of labeled DNA probes to denatured DNA fibers, and the immunodetection of the probes. We end the protocol discussion by addressing some of the more frequently asked questions with regard to common problems encountered and the technical limitations of the procedure.

Original languageEnglish (US)
Title of host publicationPlant Cytogenetics
Subtitle of host publicationGenome Structure and Chromosome Function
PublisherSpringer New York
Pages287-293
Number of pages7
Volume9
ISBN (Electronic)9780387708690
ISBN (Print)9780387708683
DOIs
StatePublished - Jan 1 2012

Keywords

  • Fiber fluorescent in situ hybridization
  • Fiber-FISH
  • Immunostaining
  • Optical mapping
  • Organelle DNA mapping
  • Repetitive DNA mapping

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