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KMID : 0613820160260091007
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Journal of Life Science
2016 Volume.26 No. 9 p.1007 ~ p.1014
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Rapid and Specific Identification of Genus Cynoglossus by Multiplex PCR Assays Using Species-specific Derived from the COI Region
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Noh Eun-Soo
Kang Hyun-Sook
An Cheul-Min
Park Jung-Youn
Kim Eun-Mi
Kang Jung-Ha
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Abstract
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A highly efficient, rapid, and reliable multiplex polymerase chain reaction based method for distinguishing ten species of genus Cynoglossus (C. senegalensis, C. abbreviates, C. macrolepidotus, C. arel, C. semilaevis, C. interruptus, C. joyneri, C. lingua, C. robustus, and C. monodi) is described. The species-specific primer sets were designed base on the cytochrome oxidase subunit I gene (1,500 bp). The optimal PCR conditions and primers were selected for ten of Cynoglossus species to determine target base sequences using single PCR. Multiplex PCR using the ten pairs of primers either specifically amplified a DNA fragment of a unique size or failed, depending on each species DNA. The length of amplification fragment of 208 bp for C. senegalensis, 322 bp for C. abbreviates, 493 bp for C. macrolepidotus, 754 bp for C. arel, 874 bp for C. semilaevis, 952 bp for C. interruptus, 1,084 bp for C. joyneri, 1,198 bp for C. lingua, 1,307 bp for C. robustus, and 1,483 bp for C. monodi with the species-specific primers, visualized by agarose gel electrophoresis, allowed perfectly distinction of the Cynoglossus species. The multiplex PCR assay can be easily performed on multiple samples and attain final results in less than 6 hours. This technique should be a useful addition to the molecular typing tools for the tentative identification of Cynoglossus species.
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KEYWORD
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Cynoglossus spp., mitochondrial DNA, molecular identification, multiplex PCR, species-specific primer
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