Development and Validation of PCR-RFLP Assay to Identify Three Japanese Brown Frogs of the True Frog Genus Rana

DOI Web Site 被引用文献4件 参考文献6件 オープンアクセス
  • IGAWA Takeshi
    Division of Developmental Science, Graduate School of International Development and Cooperation, Hiroshima University
  • KOMAKI Shohei
    Institute for Amphibian Biology, Graduate School of Science, Hiroshima University
  • TAKAHARA Teruhiko
    Laboratory of Forest Ecology, Graduate School of Integrated Arts and Sciences, Hiroshima University
  • SUMIDA Masayuki
    Institute for Amphibian Biology, Graduate School of Science, Hiroshima University

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タイトル別名
  • Development and Validation of PCR-RFLP Assay to Identify Three Japanese Brown Frogs of the True Frog Genus <i>Rana</i>
  • Development and validation of PCR-RFLP assay to identify three brown frogs of the true frog genus Rana

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Two species of the true frog genus Rana (R. japonica and R. ornativentris) are widely distributed on the main islands of Japan with some degree of habitat overlap. These species could be key indicators for environmental conditions and accurate identification is important. However, morphological characters in early stages of development are often insufficient to distinguish the two species, although their presence is often inferred from egg masses. Therefore, we developed a PCR-RFLP protocol for efficient and cost-effective identification of the species. We used a short partial 16S rRNA fragment (approx. 550 bp) and two diagnostic restriction enzymes (Spe I and Hph I) to identify species from DNA material. This approach allowed us to accurately assign all samples even without locality data. We also confirmed the performance of our method using another brown frog, R. t. tagoi.

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