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Molecular identification of species and ploidy of Carassius fishes in Lake Biwa,using mtDNA and microsatellite multiplex PCRs |
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| Authors: | Tappei Mishina Mikumi Takada Hirohiko Takeshima Mitsunori Nakano Ryoichi Tabata Mutsumi Nishida Katsutoshi Watanabe |
| Institution: | 1. Laboratory of Animal Ecology, Department of Zoology, Division of Biological Science, Graduate School of Science, Kyoto University, Kitashirakawa-Oiwake-cho, Sakyo-ku, Kyoto, 606-8502, Japan 2. Atmosphere and Ocean Research Institute, The University of Tokyo, Kashiwanoha, Kashiwa-shi, Chiba, 277-8564, Japan 3. Environmental Science Graduate School, The University of Shiga Prefecture, 2500 Hassaka-cho, Hikone-City, Shiga, 522-8533, Japan 4. University of the Ryukyus, 1 Senbaru, Nishihara-cho, Nakagami-gun, Okinawa, 903-0213, Japan |
| Abstract: | We established an efficient and reliable method to identify the species and ploidy of Carassius fishes in Lake Biwa (C. cuvieri, C. buergeri, and the latter’s polyploid) using a combination of multiplex mitochondrial DNA haplotype-specific PCR and ploidy determination based on nuclear DNA microsatellite PCR. The high accuracy of ploidy determination was confirmed by flow cytometry. This method can also identify triploid clonal lines and provide information about their genetic diversity and structure. The method is applicable to ecological and evolutionary studies of Carassius fishes in Lake Biwa throughout their entire life history. |
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