Application of PCR-RFLP analysis on species identification of canned tuna

Abstract

The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was developed to identify the species of Thunnus thynnus, T. alalunga, T. obesus, T. albacares, Euthynnus pelamis (Katsuwonus pelamis), E. affinis, Auxis thazard and Sarda orientalis in products of canned tuna. To increase the efficiency and sensitivity of DNA extraction, a recent developed method of binding magnetic beads was applied in this study. Two sets of primer were designed to amplify 126 bp and 146 bp of partial mitochondrial cytochrome b gene, and five restriction enzymes including Bsp1286I, HincII, RsaI, ScaI and MboII were determined to analyze the short length fragments. The developed PCR-RFLP method was applied to authenticate species of 18 commercial canned tuna successfully, and it really provided a useful and academic technique to identify eight species in canned tuna.