Abstract
A unique DNA isolation protocol based on microwave irradiation and density centrifugation for the simultaneous detection of porcine, chicken, and mutton meat species in food by PCR using newly developed primers is described. Emphasis is placed on the detection of pork, as it is considered unlawful in the diet in major religions such as Islam and Judaism. The method has been optimized using the primers designed from porcine mitochondrial DNA (mtDNA) sequence and tested in 96 samples. The method involves 35 seconds of high microwave irradiation of lysed sample homogenate followed by a 5- min centrifugation in 1.5 mL Eppendorf tubes until a clear supernatant is obtained. The design of this protocol makes it possible to process many samples in a short time. To evaluate the method pork and food spiked with different concentrations of pork together with unknown food samples were analyzed by PCR using the new set of porcine specific primers. The new set of primers and the method involved showed high sensitivity in detecting porcine and chicken content in food in addition as a marker for the detection of mutton. The present sample preparation method has the potential to be applied to other meat detection systems as well.
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