Mammals can be molecularly sexed by polymerase chain reaction (PCR) amplification of Y chromosome fragments or coamplification of homologous fragments from both sex chromosomes, which are discriminated by size polymorphism or Y-specific restriction digestion. Although coamplification of X and Y fragments is more reliable, size polymorphism in homologous fragments is uncommon and Y-specific restriction site identification requires screening with a battery of enzymes or cloning. Here we describe a simple approach, using 'double peaks' in the chromatogram upon direct sequencing of PCR products from males, to identify Y-specific restriction sites, and demonstrate its utility by application to a range of taxa. We demonstrate its utility by application to a range of mammalian taxa; Probocideans: Asian elephant (Elephas maximus), Perrisodactyls: Indian rhinoceros (Rhinoceros unicornis), Carnivores: domestic dog (Canis familiaris) and Primates: Tonkean macaque (Macaca tonkeana).
Molecular Ecology Notes 1, 350-353.