Background: An unidentified animal species named the Jenglot and claimed to be a rare living animal species was recently found in the deep jungle of Irian Jaya, Indonesia; brought to Kuala Lumpur, Malaysia by a businessman; and exhibited in a local museum. The owner of the Jenglot carcasses had made a request to perform DNA analysis on the Jenglot to ascertain its species. Methods: Because the muscle appeared very dry and recovery of DNA was extremely difficult, we therefore used the animals’ hair for further analysis. Hair samples were collected from three different Jenglots that were different in colour and physical appearance. The samples were labelled as A, B, C and D, respectively. Results: Microscopic characteristics indicated that all four hair samples were of human origin, with a medullary index less than 1/3 and pigment distribution towards the periphery. The scale pattern on the hair samples was of the imbricate type, adding certainty to the hypothesis of human origin. A dried root sheath was found in samples B and C, which was contrary to expectations since the sample collection method left a few cm of hair on the body of the Jenglots. Sample D had black dye granules over the cuticular surface. Sequencing of the mitochondrial DNA (mtDNA) hypervariable segment I (HVS-I) region showed polymorphisms at positions 16140, 16182C, 16183C, 16189, 16217 and 16274 and heteroplasmy at positions 16112, 16232 and 16251, a human-specific mtDNA haplotype that was consistent across all the samples. Conclusions: Based on these findings, it was concluded that it is unlikely that the samples of Jenglot hair originated from an animal species.