Gene mapping by chromosome spot hybridization

J. G. Collard*, P. A. J. De Boer, J. W. G. Janssen, J. F. Schijven, B. De Jong

*Corresponding author for this work

    Research output: Contribution to journalArticleAcademicpeer-review

    20 Citations (Scopus)

    Abstract

    A method is described for the localization of cloned single-copy genes to flow-sorted chromosomes. Chromosomes were sorted directly onto nitrocellulose filters and the chromosomal DNA was subsequently hybridized with gene-specific radioactively labeled DNA probes. Mild aspiration of the filters during sorting was applied to collect the deflected chromosomes in a small spot. Sorting of 10,000-30,000 chromosomes was sufficient to detect gene-specific hybridization with single-copy DNA probes. Using this technique, we have sublocalized the human c-myb oncogene to 6q21-q23 by sorting translocated chromosomes with breakpoints in the q21 and q23 region of chromosome 6. Chromosome spot hybridization appears to be a rapid and simple method to assign cloned genes to chromosomes. Hybridization of an unlocalized gene probe to spots of chromosomes pre-enriched by velocity sedimentation can quickly narrow the choice of chromosomes which need to be sorted. Conversely, individual chromosomes in a flow karyotype can be identified by hybridizing sorted chromosomal DNA with chromosome-specific DNA probes.
    Original languageEnglish
    Pages (from-to)179-185
    Number of pages7
    JournalCytometry
    Volume6
    Issue number3
    DOIs
    Publication statusPublished - 1985

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