In natural history collections throughout Europe, there are many old lion specimens of unknown origin. If these specimens can be shown to have originated from now-extinct populations their value would significantly increase, as would the value of the collections. Recently, a 200-year old mounted skeleton in the Zoological Museum Amsterdam has been identified as the extinct Cape lion Panthera leo melanochaita (Smith, 1842), based primarily on morphological information inferred from a painting of this specimen while it was still alive. To test this hypothesis, we used ancient DNA (aDNA) techniques to extract and sequence mitochondrial DNA (mtDNA) from this specimen, and compared the genetic results with previously published lion mtDNA sequences. Our results show that the specimen is not a Cape lion, but that it instead possesses the mtDNA haplotype of the Asiatic lions P. l. persica (Meyer, 1826) from India. This Indian origin hypothesis is further supported by an investigation of its cranial morphology. As the amount of genetic information available for lions increases, in particular data from across their historic distribution, the potential for aDNA techniques to identify the origins of previously unassigned museum specimens continues to grow.