This study describes an investigation into the use of a PCR-RFLP technique as a routine analytical tool for species testing. The technique was used to generate DNA fingerprints for 22 animal species by amplifying a 359 bp region within the cytochrome b gene and digesting the amplified product using Hae III and Hinf I. All species could be discriminated using the two restriction enzymes with the exception of kangaroo and buffalo. Cooking the tissues did not affect the DNA extractions or the profiles generated. When mixtures were investigated, pig was preferentially amplified and dominated over all species tested, even at levels of 1%. Another set of cytochrome b primers which amplified 464 bp, was also tested for the analysis of these mixtures. Beef was found to be favourably amplified over the other species. Anomolous results where the digested products exceeded 359 bp was also investigated. Co-amplification was found to occur in the species investigated. Results of this study suggest that the CytB PCR-RFLP method shows promise for the identification of both cooked and uncooked tissues, although the method is unsuitable for analysing meat mixtures.
Meat Science – Elsevier
Published: Apr 1, 2000
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