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The electronic form of this document may be cited in the following style:
Human Genome Program, U.S. Department of Energy, DOE Human Genome Program Contractor-Grantee Workshop IV, 1994.

Abstracts scanned from text submitted for November 1994 DOE Human Genome Program Contractor-Grantee Workshop. Inaccuracies have not been corrected.

Generating and Typing CAPS Genetic Markers in Dog

Mark W. Neff[1,2], Mike Strathmann[1], and Jasper Rine[2 ]
[1]Human Genome Center, Lawrence Berkeley Laboratory, Berkeley, CA 94720. [2]Department of Molecular and Cellular Biology, University of California, Berkeley, CA 94720.
EM: Neff@Mendel.berkeley.edu.

The behavioral and morphological traits that distinguish the various dog breeds are genetically determined as they have been fixed by artificial selection. A genetic map of the dog genome will make breed-specific traits amenable to quantitative trait loci (QTL) analysis. Presently, a genetic map of dog consisting of microsatellite markers is being assembled [1]. Although polymorphic and informative, these molecular markers have several genotyping limitations. They require polyacrylamide gel electrophoresis, radioactive- or fluorescence-based detection, and manual scoring. Furthermore, only a few markers can be typed per reaction such that genotyping efforts increase linearly with map resolution. For a quantitative genetics approach involving controlled matings, a less polymorphic, more easily typed marker might also be valuable.

A cleaved amplified polymorphic sequence (CAPS) is a RFLP encompassed by primer sites [2]. Although less polymorphic than microsatellite markers, CAPS markers may be typed simply by agarose electrophoresis and ethidium bromide staining. Genomic subtraction methods have previously been used to generate libraries of RFLP loci [3]. We report on the construction of a CAPS library based on an experimental Border Collie X Newfoundland cross. We also present progress on a novel method for genotyping many CAPS markers simultaneously. These advances should reduce the resources and effort necessary to construct and use genetic maps.

M.N. was supported by a DOE Human Genome Postdoctoral Fellowship, and M.S. was supported by an Alexander Hollander Postdoctoral Fellowship.

[1] E. A. Ostrander, G. F. Sprague and J. Rine (1993) Identification and characterization of dinucleotide repeat (CA)n markers for genetic mapping in dog. Genomics 16: 207-213.
[2] A. Konieczny and F. M. Ausubel (1993) A procedure for mapping Arabidopsis mutations using co-dominant ecotype-specific PCR-based markers. Plant J. 4: 403-10.
[3] M. Rosenberg, M. Przybylska and D. Straus (1994) "RFLP subtraction": a method for making libraries of polymorphic markers. Proc. Natl. Acad. Sci. USA 91: 6113-7.