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Tigers are killed to supply a demand for many wildlife products despite a ban on commercial international trade. As populations decrease, products from substitute species (i.e. lions and leopards) have been fraudulently sold as tiger. DNA forensic techniques are needed to definitively identify tiger in order to secure prosecutions although this is complicated by the presence of numts. Therefore, we have developed and validated a CO1 genetic marker that preferentially amplifies the mtDNA CO1 region and excludes the nuclear CO1 pseudogene, which we expect to be of use in tiger forensic casework.