The human chromosome content in human x rodent somatic cell hybrids analyzed by a screening technique using Alu PCR.
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.
Bicknell
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DC
Cancer Immunology Laboratory, University of Oxford, John Radcliffe Hospital, Headington, United Kingdom.
Markie
D
D
Spurr
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NK
Bodmer
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WF
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
D004247:DNA