Phylogenetic analysis of organisms not only offers crucial details on the genesis and evolution of genes, genomes, and species, but also helps with ecological and behavioural research while forecasting the magnitude and direction of future evolutionary trends in living things. In the present study, cytochrome b gene sequence of mitochondrial DNA obtained from captive Bengal tiger (Panthera tigris) from the Arignar Anna Zoological Park, Vandalur, was phylogenetically analysed with reference sequences of cytochrome b gene of fourteen sequences of the genus Panthera obtained from NCBI gen bank database. The similarities and differences between the different sequences were studied and the use of phylogenetic analysis as a forensic tool for investigation was discussed. The phylogenetic tool can also be used for pedigree checking where the parentage details are maintained and the place of origin of the species can be traced in case of samples seized during animal poaching.