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Updated: Jun 23, 2025

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High-Resolution FISH Analysis Using DNA Fibers Generated by Molecular Combing.

Sandra Louzada1,2, Fengtang Yang3

  • 1CytoGenomics Lab, Department of Genetics and Biotechnology (DGB), University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal.

Methods in Molecular Biology (Clifton, N.J.)
|June 24, 2024
PubMed
Summary

Molecular combing stretches DNA molecules for high-resolution imaging. Combining it with fluorescent in situ hybridization (FISH) allows detailed visualization of DNA structure and variations.

Keywords:
DNA fibersFluorescence in situ hybridizationMolecular combingMulticolor fiber-FISH

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Area of Science:

  • Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • Molecular combing is a DNA stretching technique offering high resolution.
  • Fluorescent in situ hybridization (FISH) allows DNA visualization.
  • Combining these methods enhances DNA structure analysis.

Purpose of the Study:

  • To detail the molecular combing procedure for DNA fiber preparation.
  • To demonstrate applications of molecular combing combined with multicolor fiber-FISH.
  • To highlight the technique's utility in high-resolution genomic studies.

Main Methods:

  • DNA fiber preparation using molecular combing on a glass surface.
  • Application of multicolor fluorescent in situ hybridization (FISH) to combed DNA.
  • High-resolution imaging and analysis of DNA structures.

Main Results:

  • Achieved a resolution of two kilo-basepairs per micrometer.
  • Successfully visualized DNA structure and variations at unprecedented resolution.
  • Demonstrated utility in identifying copy number variations and structural variations.
  • Enabled precise measurement of overlaps and gaps in genome assemblies.

Conclusions:

  • Molecular combing coupled with FISH provides a powerful tool for high-resolution DNA analysis.
  • The technique facilitates detailed examination of genomic structure and variations.
  • It offers significant advantages for genome assembly and structural variation studies.