The objective of this study was to obtain a fast, accurate and reliable method of species identification of unknown biological samples for forensic applications, especially in illegal trade of animals as well as meat fraud. Meat fraud and adulteration not only affects the market but also increases the risk of religious and ethnic conflicts around the world [1]. In this study, species-specific and gender differentiating Real time PCR technique was employed to analyse 15 meat samples collected from a suspected site. Out of 15 samples collected from suspected site, 54% and 13% samples were of Cow and buffalo origin respectively. All 54% cow samples were of male while one each of buffalo were of male and female origin. Two samples were inconclusive. These findings indicated that species and gender-specific PCR is very sensitive and can be used for forensic species identification and the detection of meat fraud and adulteration.