We have developed a species identification method from environmental DNA using inexpensive PCR-RFLP. (Joint research with Prof. Takahara, Shimane Univ.)

Although environmental DNA has now become a standard tool in ecological research, it requires expensive equipment and reagents such as real-time PCR instruments and fluorescent probes, and the laboratories that can prepare them are limited.

Therefore, we have developed a method for efficiently detecting multiple species simultaneously and inexpensively from environmental water by applying the PCR-RFLP (PCR product restriction enzyme fragment length polymorphism) method. This method does not require a real-time PCR device or fluorescently labeled oligo DNA, and can be used for experiments only with a PCR device and an electrophoresis device in a general laboratory. It can be implemented even in laboratories mainly for ecological research, and some junior high and high schools with PCR devices.

The target species, the Japanese brown frog (Rana japonica), the mountain brown frog (Rana ornativentris), and the Tago frog (Rana tagoi tagoi), are common amphibians that are often found in rural and mountain villages of Japan and are also key indicators for environmental monitoring. I hope that the method developed this time will help build a sustainable society in harmony with nature.

Reference：*Igawa T, *Takahara T, Lau Q, Komaki S, An application of PCR-RFLP species identification assay for environmental DNA detection. PeerJ 7: e7597 October 2019. [ DOI | http | PDF ]
*co-corresponding authors