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Related Experiment Videos

Visual mapping by fiber-FISH

M Heiskanen1, E Hellsten, O P Kallioniemi

  • 1Department of Clinical Chemistry, University of Helsinki, Finland.

Genomics
|November 1, 1995
PubMed
Summary
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Improved fiber-FISH offers high-resolution physical mapping and contig construction. This technique accurately orders DNA probes and detects genomic rearrangements, aiding positional cloning and disease gene discovery.

Area of Science:

  • Genomics
  • Molecular Biology
  • Cytogenetics

Background:

  • Fluorescence In Situ Hybridization (FISH) techniques are crucial for high-resolution genome mapping.
  • Accurate ordering and characterization of genomic contigs are essential for physical mapping and positional cloning.
  • Existing methods require improvement for efficient analysis of large genomic regions.

Purpose of the Study:

  • To evaluate an improved fiber-FISH technique for high-resolution physical mapping.
  • To assess its utility in characterizing a 500-kb genomic region at 1p32.
  • To demonstrate its capability in detecting genomic rearrangements.

Main Methods:

  • Developed and utilized an improved fiber-FISH technique involving hybridization to unfixed, linearized DNA fibers.

Related Experiment Videos

  • Prepared target DNA from cells embedded in pulsed-field gel electrophoresis (PFGE) blocks for long, intact DNA fibers.
  • Applied the technique to order and orient multiple probes (phage, cosmid, P1 clones, subclones) and detect rearrangements.

    • Main Results:

    • The improved fiber-FISH technique achieved a linear range from 2 to 500 kb with high measuring accuracy.
    • Successfully established probe order, orientation, and distances for a 500-kb region at 1p32.
    • Demonstrated the visualization of genomic rearrangements, including those affecting the L-myc gene in small-cell lung cancer.

    Conclusions:

    • Fiber-FISH is highly effective for high-resolution physical mapping and genomic contig evaluation.
    • The technique facilitates positional cloning efforts, including the identification of disease genes like INCL.
    • Fiber-FISH is a valuable tool for detecting and characterizing genetic rearrangements.