Gene mapping by chromosome spot hybridization.
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.
D002458:Cell Fractionation; D002478:Cells, Cultured; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005347:Fibroblasts; D005434:Flow Cytometry; D005796:Genes; D006801:Humans; D009693:Nucleic Acid Hybridization; D009857:Oncogenes
D004247:DNA