Identification of Thunnus Species by PCR-RFLP Method

This technical report describes the identification of various tuna species using the PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) method, facilitated by the MultiNA II microchip electrophoresis system. The document outlines the procedures for DNA extraction from different tuna species, including Atlantic bluefin tuna, southern bluefin tuna, α and β-type bigeye tuna, yellowfin tuna, and albacore tuna. It details the PCR process with specific primers for mitochondrial DNA and the subsequent restriction enzyme digestion. The report presents the results of electrophoresis patterns that distinguish these fish varieties based on their DNA fragmentation patterns after processing with enzymes such as Alu I, Mse I, and Tsp509 I. The findings indicate that the MultiNA II system allows for automated analysis of samples, enhancing the accuracy and efficiency of species identification, which is crucial for compliance with Japan's food labeling standards. This method addresses the challenges of misidentification and mislabeling in seafood distribution.