A technique to discriminate Canis mitochondrial DNA of new world and old world origins using specific primers

Genetic markers play an important role in elucidating taxonomic uncertainties for a wide range of organisms. We present a set of specific primers to distinguish between Canis mitochondrial DNA (mtDNA) of New World (North American) and Old World (Eurasian) origin using the ATP-8 region and gel electrophoresis. We amplified mtDNA from Old World (gray wolves [Canis lupus L., 1758]) and New World canids (coyotes [C. latrans Say, 1823] and eastern wolves [C. lycaon Schreber, 1775 or C. lupus lycaon]) collected during 2003-2009 in Québec, Canada, using a multiplexed primer triplet. The results showed a standard band of 150 base pairs (bp) for New World and Old World mtDNA. In addition, Old WorldmtDNA displayed a second band of 100 bp. The range extent of wolves with New World mtDNAhas important implications for canid conservation. The new method can assist conservation managers with rapid and cost-effective screening to monitor 1) the distribution and abundance of wolves with New World and Old World mtDNA, and 2) wolf-coyote hybridization, when used in combination with morphological information and other nuclear markers.