ABSTRACT
Abstract: Today's, consumers are concerned about the meat they eat and also demandaccurate labeling. Mitochondrial Analysis of DNA was the most frequently used DNA, becauseof its highly conserved sequences in various organism species. In this study, a rapid,reproducible and simple method for simultaneous identification of multiple meat species ina single step mitochondrial DNA based test has been developed based on the designing ofspecies-specific primer. Meat samples of goat and buffalo were selected to verify theapplicability of the technique. A species specific forward and reverse primer was designedwith the help of the primer3 tool for amplification of mitochondrial D-loop region. Thespecies-specific primers were verified in silico by SnapGene software. The two pairs ofprimers amplified the expected fragment of 338bp for buffalo and 450bp for goat. The changein the size of the PCR product was due to the existence of highly polymorphic regions withinthe buffalo and goat D-loop region. The tested species gives a unique band pattern for eachspecies by using successful amplification of these polymorphic regions in the D-loop region.Overall, the simplicity of amplification of mitochondrial D-loop region could make thistechnique suitable for meat authentication in routine analysis.