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A method for constructing radiation hybrid maps of whole genomes: Application to physically mapping chromosome 14

Journal Article · · American Journal of Human Genetics
DOI:https://doi.org/10.1038/ng0594-22· OSTI ID:134489
;  [1];  [2]
  1. Univ. of Alberta (Canada)
  2. Niigata Univ. (Japan); and others

By reverting to the original protocols of Goss and Harris, we have created a panel of whole genome radiation hybrids (WG-RHs) using a diploid human fibroblast as the chromosome donor, rather than the usual monochromosomal human/rodent somatic cell hybrid. We have analyzed markers from chromosome 14 to test the feasibility of using WG-RH cell lines to generate physical maps of human chromosomes. As WG-RH mapping exploits rodent/human differences, loci need not be polymorphic to be informative. Sixty-one chromosome 14 markers, including 24 STSs and ETSs, were used to create a high resolution radiation hybrid map of human chromosome 14. The average marker retention was found to be 22.4%, very similar to the marker retention frequencies of conventional radiation hybrids. Two point and multipoint statistical analyses of the patterns of chromosome 14 marker retention were used to create a WG-RH map of human chromosome 14 with 4 gaps, corresponding to regions of low marker density. We are currently testing additional markers to close the map. Conventional radiation hybrid mapping requires between 100 and 200 hybrids to map each chromosome. The large number of hybrids (up to 4,000) required to map the whole genome is a major drawback of this method. In contrast, a single panel of 100 to 200 WG-RH cell lines is sufficient to allow the construction of a high resolution map of the whole human genome with a single panel of only 100 to 200 hybrids. Our results demonstrate that chromosome fragmentation by WG-RH can be used to map one chromosome, and by extension, entire genomes.

Sponsoring Organization:
USDOE
OSTI ID:
134489
Report Number(s):
CONF-941009--
Journal Information:
American Journal of Human Genetics, Journal Name: American Journal of Human Genetics Journal Issue: Suppl.3 Vol. 55; ISSN AJHGAG; ISSN 0002-9297
Country of Publication:
United States
Language:
English

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