Real-time multiplex PCR for simultaneous detection of multiple species from environmental DNA: an application on two Japanese medaka species

• DOI: 10.1038/s41598-018-27434-w
• Publication Date: 2018-06-08T10:43:44Z
• Authors: Satsuki Tsuji; Yuka Iguchi; Naoki Shibata; Iori Teramura; Tadao Kitagawa; Hiroki Yamanaka
• Keywords: Environmental DNA; Oryzias; Multiplex; Primer (cosmetics)

Abstract Information about species distribution is crucial to ecological studies. Environmental DNA (eDNA) analysis has recently been used estimate the of aquatic organisms. Several analytical methods including metabarcoding and species-specific PCR are being for eDNA analysis. However, when only a few targeted, not cost-effective because wasted consumption read due amplification non-target DNA. On other hand, requires tests be repeated multiple times resulting in consuming more templates, experimental consumables. Here we propose methodological framework simultaneously detecting using real-time multiplex PCR. We developed primer-probe sets two Japanese medaka ( Oryzias latipes o. sakaizumii ), them In aquarium experiment, even abundances were biased, both detected all samples. field survey, capture survey produced consistent results sampling sites, sites with low fish densities. can easily applied organisms, enabling target