title: The human chromosome content in human x rodent somatic cell hybrids analyzed by a screening technique using Alu PCR.
abstract: The ubiquitous nature of the Alu sequence throughout the human genome forms the basis of an assay we present here for analyzing the human chromosome content of human x rodent somatic cell hybrids. A human-specific Alu primer was used both to amplify sequences and to 32P label the products in a polymerase chain reaction (PCR) technique. Unlabeled inter-Alu PCR products from two series of human x rodent hybrids were used to prepare dot blots which were probed with labeled inter-Alu products prepared from between 10(3) and 10(4) hybrid cells. In the first series we demonstrate that a labeled inter-Alu probe from the hybrid DL18ts, containing a single chromosome 18, on a dot blot hybridized only with those inter-Alu products containing chromosome 18. Similar specificity for human chromosome 5 was shown when a Southern blot of the PCR products was hybridized with a probe made from the hybrid HHW 213, which contains only chromosome 5p. Using a dot blot from a second series of control hybrids, 15 of which contained single human chromosomes, hybridization of a labeled probe from the hybrid 18X4-1 was shown to react specifically with the controls that expressed chromosome 18. Application of the technique reported here allows simple and rapid characterization of the human chromosome content in human x rodent hybrids.
authors.affiliation: Cancer Immunology Laboratory, University of Oxford, John Radcliffe Hospital, Headington, United Kingdom.
authors.forename: D C
authors.initials: DC
authors.lastname: Bicknell
authors.forename: D
authors.initials: D
authors.lastname: Markie
authors.forename: N K
authors.initials: NK
authors.lastname: Spurr
authors.forename: W F
authors.initials: WF
authors.lastname: Bodmer
mesh_terms: D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002877:Chromosomes, Human; D006224:Cricetinae; D004247:DNA / Q000302:isolation & purification; D006801:Humans; D006822:Hybrid Cells; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D013698:Templates, Genetic
chemical_list: D004247:DNA