Effects of preservation strategies on environmental DNA detection and quantification using ddPCR

Molecular-based monitoring relying on environmental DNA (eDNA) detection became routinely used around the world in last few years, especially aquatic environments. The large potential and increasing applications of this technique calls for technical improvements to optimize reliability these surveys. An important aspect eDNA workflow is appropriate preservation samples taken field, as it can significantly affect recovery ultimately false negative rates. In study, we explored efficiency five different strategies by using a controlled mesocosm experiment which included three fish communities composition. Specifically, compared extractions (a) performed immediately following collection, or after eight months storage from (b) frozen filters, (c) unfiltered water stored at ?20°C, filters preserved room temperature with (d) Longmire (e) Sarkosyl buffer. Effects were quantified ddPCR measurements species (Neogobius melanostomus, Rutilus rutilus, Lota lota) total content group-specific primers Teleostei. Samples extracted collection without any further yielded less other approaches. Overall, Longmire's buffer facilitated best across all although approaches such filter freezing use similar results. Relative measurement variability, an indicator reliable quantification, was lowest when buffers generally decreased quantity. our results clearly highlight significant impact sample how substantially performance eDNA-based