The goal in my study is to develop a set of universal genus-spesific PCR primers for eDNA metabarcoding in order to correctly distinguish the eDNA of all the 19 species and subspecies of the genus Anguilla from water samples. To examine the perform of the newly designed primers, we collected eDNA samples form mock communities, tank waters with known species compositions and natural rivers in Japan, and then prepared dual-indexed libraries and performed paired-end sequencing with high-throughput next-generation sequencing (Illumina, MiSeq) technologies. The reads with identify of more than or equal to 97% on MiSeq sequencing are assigned to each correct species using the custom pipeline. We can detect eDNA of each all the genus Anguilla from the water samples, suggesting our designed primers provide us the presence, absence and distribution of their genus in aquatic environments.