A PCR- RELP tool for differentiating cattle, buffalo and nilgai

Abstract

Species identification and differentiation is essential to prevent illegal trade in fauna and products. In this study, PCR-RFLP was used to differentiate nilgai (Boselaphus tragocalnelus) from its closely related domestic counterparts cattle and buffalo using commonly available restriction enzymes (Dde 1, Hinf I and Hha I). Genomic DNA was isolated from blood of these species. The 440bp region of 12S rRNA gene was amplified using universal primers and subjected to restriction digestion. Dde I digestion yielded the RFLP pattern which distinguished all three species whereas other two enzymes HinfI and Hha I were specific for cattle and buffalo respectively. This molecular method could prove a useful tool in forensic identification of cattle.