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http://dx.doi.org/10.5352/JLS.2018.28.9.1062

Development of the Duplex PCR Method of Identifying Trachurus japonicus and Trachurus novaezelandiae  

Park, Yeon Jung (Biotechnology Research Division, National Institute of Fisheries Science)
Lee, Mi Nan (Biotechnology Research Division, National Institute of Fisheries Science)
Kim, Eun Mi (Biotechnology Research Division, National Institute of Fisheries Science)
Noh, Eun Soo (Biotechnology Research Division, National Institute of Fisheries Science)
Noh, Jae Koo (Biotechnology Research Division, National Institute of Fisheries Science)
Park, Jung Youn (Biotechnology Research Division, National Institute of Fisheries Science)
Kang, Jung-Ha (Biotechnology Research Division, National Institute of Fisheries Science)
Publication Information
Journal of Life Science / v.28, no.9, 2018 , pp. 1062-1067 More about this Journal
Abstract
Reliable labeling of fish products can reassure consumers regarding the identity and quality of seafoods. Therefore, techniques that can identify adulteration or mislabeling are valuable. To rapidly identify two Trachurus species, Trachurus japonicus and Trachurus novaezelandiae, a highly efficient, rapid, duplex polymerase chain reaction (PCR) having two species-specific primers simultaneously was identified. This species-specific primer focused on a single nucleotide mismatch in the 3'-terminal base of a primer designed in the mitochondrial cytochrome c oxidase (COI) subunit I DNA. To optimize the duplex PCR condition, gradient PCR reactions were conducted to determine the primer annealing temperature and the primer concentration. The PCR's product was observed on the gel, suggesting that DNA molecules may be useful in differentiating the two species. The length of the amplification fragments were 103 bp for Trachurus japonicus and 214 bp for Trachurus novaezelandiae, which, along with the species-specific primer visualized by agarose gel electrophoresis, enabled accurate distinction of the species of the Trachurus genus. These results indicate that the duplex PCR, which has a species-specific primer based on single nucleotide polymorphism (SNP), can be useful for rapidly differentiating the two species of Trachurus. This duplex PCR analysis is simple, rapid, and reliable, and could be beneficial to protecting consumers' rights.
Keywords
Genus Trachurus; mitochondrial DNA; multiplex PCR; species specific primer;
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