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Related Experiment Videos

Fiber-FISH: experiences and a refined protocol

M Heiskanen1, O Kallioniemi, A Palotie

  • 1Laboratory Department of Helsinki, University Central Hospital, Finland.

Genetic Analysis : Biomolecular Engineering
|March 1, 1996
PubMed
Summary
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We refined a Fiber-FISH technique using agarose-embedded DNA for faster physical mapping and genomic rearrangement detection. This method improves the visualization of large insert clones, aiding positional cloning efforts.

Area of Science:

  • Genomics
  • Molecular Biology
  • Cytogenetics

Background:

  • Positional cloning relies heavily on physical mapping of probes and assembling genomic contigs, which is often time-consuming.
  • High-resolution Fiber-FISH techniques have improved the visualization of large insert clones, aiding in order, overlap, and gap size determination.

Purpose of the Study:

  • To present a refined protocol for a modified Fiber-FISH technique using agarose-embedded DNA.
  • To demonstrate the application of this technique for physical mapping and detecting genomic rearrangements.
  • To address critical steps and troubleshooting for the method.

Main Methods:

  • Developed a method utilizing agarose-embedded DNA (PFGE block) as a source for linearized DNA targets on microscope slides.
  • Modified the Fiber-FISH technique for enhanced visualization of large insert clones.

Related Experiment Videos

  • Applied the technique to agarose-embedded yeast cells.

    • Main Results:

    • Successfully used the modified Fiber-FISH technique for physical mapping in the 1-300 kb range.
    • Demonstrated the utility of the method for detecting genomic rearrangements.
    • Provided a refined protocol with troubleshooting guidance.

    Conclusions:

    • The refined agarose-embedded Fiber-FISH technique offers an efficient approach for physical mapping and genomic rearrangement analysis.
    • This method facilitates the assembly of genomic contigs and aids in positional cloning.
    • The protocol is applicable to various biological samples, including yeast cells.