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Proceedings Paper

Genetic linkage analysis using pooled DNA and infrared detection of tailed STRP primer patterns
Author(s): William S. Oetting; Scott C. Wildenberg; Richard A. King
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Paper Abstract

The mapping of a disease locus to a specific chromosomal region is an important step in the eventual isolation and analysis of a disease causing gene. Conventional mapping methods analyze large multiplex families and/or smaller nuclear families to find linkage between the disease and a chromosome marker that maps to a known chromosomal region. This analysis is time consuming and tedious, typically requiring the determination of 30,000 genotypes or more. For appropriate populations, we have instead utilized pooled DNA samples for gene mapping which greatly reduces the amount of time necessary for an initial chromosomal screen. This technique assumes a common founder for the disease locus of interest and searches for a region of a chromosome shared between affected individuals. Our analysis involves the PCR amplification of short tandem repeat polymorphisms (STRP) to detect these shared regions. In order to reduce the cost of genotyping, we have designed unlabeled tailed PCR primers which, when combined with a labeled universal primer, provides for an alternative to synthesizing custom labeled primers. The STRP pattern is visualized with an infrared fluorescence based automated DNA sequencer and the patterns quantitated by densitometric analysis of the allele pattern. Differences in the distribution of alleles between pools of affected and unaffected individuals, including a reduction in the number of alleles in the affected pool, indicate the sharing of a region of a chromosome. We have found this method effective for markers 10 - 15 cM away from the disease locus for a recessive genetic disease.

Paper Details

Date Published: 1 April 1996
PDF: 9 pages
Proc. SPIE 2680, Ultrasensitive Biochemical Diagnostics, (1 April 1996);
Show Author Affiliations
William S. Oetting, Univ. of Minnesota/Minneapolis (United States)
Scott C. Wildenberg, Univ. of Minnesota/Minneapolis (United States)
Richard A. King, Univ. of Minnesota/Minneapolis (United States)

Published in SPIE Proceedings Vol. 2680:
Ultrasensitive Biochemical Diagnostics
Gerald E. Cohn; Steven A. Soper; C. H. Winston Chen, Editor(s)

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