Improving sensitivity in environmental DNA measurement by reconcentrating DNA extracts

Environmental DNA (eDNA) methodology is widely applied in the biomonitoring of organisms, but it requires the target DNA to be detected in a simple, stable, and highly sensitive manner. Detection sensitivity of eDNA measurement becomes particularly critical when monitoring species present at low abundance. In this study, we aimed to improve the detection sensitivity through a method of DNA-extract reconcentration. This approach involves reconcentrating eDNA samples that were originally extracted using the widely adopted DNeasy Blood and Tissue Kit (Qiagen), utilizing the same kits reagents, and does not require any additional equipment or reagents. We evaluated the ability of this DNA reconcentration method using field samples including river, lake and costal marine habitats. Evaluation of this DNA reconcentration method showed that when ten conventionally extracted samples were pooled, the DNA concentration increased by approximately sevenfold, as confirmed by DNA quantification and quantitative PCR analyses, demonstrating enhanced detection sensitivity.