Human Genetics

, Volume 76, Issue 3, pp 290–292 | Cite as

A method for nucleic acid hybridization to isolated chromosomes in suspension

  • Gertrud Dudin
  • Thomas Cremer
  • Margit Schardin
  • Michael Hausmann
  • Frank Bier
  • Christoph Cremer
Original Investigations

Summary

A procedure was developed to provide differential fluorescent staining of metaphase chromosomes in suspension following nucleic acid hybridization. For this purpose metaphase chromosomes were isolated from a Chinese hamster x human hybrid cell line. After hybridization with biotinylated human genomic DNA, the human chromosomes were visualized by indirect immunofluorescence using antibodies against biotin and fluoresceine-isothiocyanate-(FITC)-labeled second antibodies. This resulted in green fluorescent human chromosomes. In contrast, Chinese hamster chromosomes revealed red fluorescent staining only when counterstained with propidium iodide. Notably, interspecies chromosomal rearrangements could be easily detected. After hybridization and fluorescent staining, chromosomes still showed a well-preserved morphology under the light microscope. We suggest that this procedure may have a useful application in flow cytometry and sorting.

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References

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Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • Gertrud Dudin
    • 1
  • Thomas Cremer
    • 2
  • Margit Schardin
    • 2
  • Michael Hausmann
    • 1
  • Frank Bier
    • 1
  • Christoph Cremer
    • 1
  1. 1.Institut für Angewandte Physik I der UniversitätHeidelberg 1Federal Republic of Germany
  2. 2.Institut für Anthropologie und Humangenetik der UniversitätHeidelberg 1Federal Republic of Germany

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