In-silico Analysis of the Cytochrome B Gene and its Role in Species Traceability from Fresh Meat Samples Through a Non-enzymatic DNA Extraction Method.

In the present study, we first performed in-silico analysis and confirmed that the mitochondrial cytochrome b gene can be used for species differentiation, as it contains both conserved and variable regions, making it suitable for species identification. Furthermore, we used the non-enzymatic DNA extraction method as a novel procedure for DNA extraction from fresh meat samples. The results revealed that 6% of the collected samples were mislabeled, in which cattle meat was substituted with buffalo meat. The Chamkani tehsil of the target area showed the highest 12% mislabeling, followed by Shah Alam tehsil at 8% and Peshawar city tehsil at 4% in the collected samples. In duplex PCR, DNA fragments of two species (Goat/Sheep, Cattle/Buffalo) were amplified, and in multiplex PCR, DNA fragments of three species (Buffalo, cattle, and horse) were amplified for the target fragments of the cytochrome B gene. The present study concluded that the non-enzymatic DNA extraction method is equally effective for meat samples. Furthermore, we also found that peri-urban areas are prone to meat mislabeling, which needs vigilance from the concerned regulatory authorities.