Plant & Animal Genome VI Conference

P71

RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD) TO DIFFERENTIATE BREEDS OF DOGS

James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850

We generated Random amplified polymorphic DNA (RAPD) fragments from DNA samples of nine different breeds of dogs to test their usefulness as breed markers, and whether they reflect a general divergence of genomes of different breeds. Ten DNA samples were pooled for each of nine dog breeds. Pools were used for Bulked Segregant Analysis with 82 decanucleotide primers in RAPD-PCR. Five primers resulted in a polymorphic fragment amplified only from one breed. Seven primers resulted in a polymorphic fragment only from two breeds, and six primers led to amplification of a fragment from three breeds. Three representative primers were tested in PCR to determine whether polymorphic fragments amplified from DNA pools are also amplified from individual DNA samples. PCR reactions with primer 372 resulted in a 600 bp fragment from six of ten Australian Shepherd and four of ten Golden Retriever samples. Primer OP P-14 resulted in amplification of a 1,800 bp fragment from five of ten German Shepherd Dog samples. Primer OP M-15 led to amplification of a 1,200 bp fragment from 20 of 21 DNA samples from seven breeds. However, the fragment was not amplified from eight of ten German Shepherd Dog or from ten Whippet samples. We conclude that RAPD fragments were specific for one or two breeds only. Comparisons of bandsharing percentages of RAPD-PCR products showed no difference within or between the nine breeds. We suggest that the identified breed-specific RAPD fragments did not reflect a general genetic divergence between breeds.