Development of Molecular method for Identificaiton of Animal Species in Meat Samples

Abstract

The present study was carried out to identify and discriminate species- specific meat viz. cow, buffalo, goat and sheep by developing multiplex PCR assay using cyt b and 12S r RNA genes variability. Meat samples randomly collected from different respective species of animals. Genomic DNA of PCR grade was extracted. The amplification of cyt b gene by conventional PCR as well as multiplex PCR, the species specific primers were designed. The primer concentration was optimized and primers were mixed in 1 :0.25:4:0.6 ratios for CFP: Goat: Buffalo: Sheep. This successfully amplified the cyt b gene from only the target species and thus produced a characteristics band pattern of155 bp for goat, 270 bp for buffalo and 330 bp for sheep respectively. The second approach was applied by simultaneous amplification of cytochrome b gene and 12S r RNA genes. The four different sets of primers as two sets of primers to amplify the 12S r RNA gene of cow and buffalo and another two set of primers to amplify the Cyt b gene of goat and sheep were desined. After testing the species specificity of each primer, a multiplex PCR was performed and amplified PCR products from four species ranged from 155 to 500 bp (Cow 500bp, Buffalo 220bp, Goat 155bp and Sheep 330bp) respectively. In conclusion, it was observed that cytochrome b gene and 12S r RNA gene could be amplified by the conventional PCR as well multiplex PCR and can be used for simultaneous detection of multiple species origin in meat and met Products