|
KMID : 0613820180280091062
|
|
Journal of Life Science
2018 Volume.28 No. 9 p.1062 ~ p.1067
|
|
|
Development of the Duplex PCR Method of Identifying Trachurus japonicus and Trachurus novaezelandiae
|
|
Park Yeon-Jung
Lee Mi-Nan
Kim Eun-Mi
Noh Eun-Soo
Noh Jae-Koo
Park Jung-Youn
Kang Jung-Ha
|
|
|
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.
|
|
|
KEYWORD
|
|
|
Genus Trachurus, mitochondrial DNA, multiplex PCR, species specific primer
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|