The Convention on the International Trade in Endangered Species of Wild Fauna and Flora (CITES) monitors the international trade in endangered animal and plant species; a high profile example is the tiger, Panthera tigris. We report on the application of a SNaPshot multiplex technique to simultaneously identify tiger species and subspecies; this test is based on identification of SNPs within the tiger mitochondrial genome. Mitochondrial DNA sequences from four of the five extant putative tiger subspecies were obtained and combined with DNA sequence data from 492 tiger and 349 other mammalian species. A total of 11 SNP loci were identified: five specific for tiger; three specific to Panthera tigris sumatrae and; three specific to P. t. tigris. The multiplex assay was able to reliably identify 15 voucher tiger samples. The sensitivity of the test was 15,000 mitochondrial DNA copies, indicating that it will work on trace amounts of tissue, bone or hair