Identification of Carnivore Animal Species by Using mt DNA PCR Technique

DNA (RAPD) fragment is confirmed by presence or absence of bands resulting from deletion, insertion or differences in the nucleotide sequence in or between priming regions (Clark and Lanigan, 1993). Some highly conserved regions on the mitochondrial (mt) cythochrome b gene are suitable for species identification in vertebrates. The PCR technique is quite reliable, accurate and authentic, but requires species-specific or conserved oligonucleotide primers for identification of species. The mt DNA PCR technique was used in the present study to identify leopard, wolf and tiger. For the present work, blood samples of leopard and wolf were collected from Maharaj Bagh Zoo, Nagpur, while the blood sample from tiger was obtained from naturally died tiger during postmortem performed at Government Polyclinic, Chandarpur. The blood samples were collected in sterile 15 ml polypropylene tube containing 0.5 ml of 0.5 M ethylene diamine tetra acetate (EDTA) solution. After collection, samples were kept at-20°C till further use. i) Isolation of DNA: DNA from tissue samples was isolated by standard methods of proteinase K digestion, phenol-chloroform extraction and DNA precipitation in ethanol (Sambrook and Russel, 2001). The DNA pellet was the