Environmental DNA (eDNA) is increasingly used as a cost-effective, non-invasive alternative or complement to traditional sampling methods, with applications ranging from targeted species detection to long-term biodiversity monitoring. The NSF-funded project, “Collaborative Research: MRA: Determining InStream Transport of eDNA across NEON sites at the Continental ScalE (DISTANCE),” examines how abiotic factors influence the transport, persistence, and fate of eDNA in stream ecosystems. We will conduct repeated, constant-rate releases of Atlantic Herring (Clupea harengus) DNA (in the form of a homogenized tissue slurry) to quantify downstream eDNA transport dynamics across National Ecological Observatory Network (NEON) stream sites spanning multiple ecoregions in the United States. These nutrient-style additions will elucidate how spiraling dynamics vary across a wide range of streams with different biogeochemical conditions. In addition to Atlantic Herring releases, we will conduct seasonal releases to characterize temporal DNA flux patterns from a suite of locally significant species with differing tissue types in North Texas within Pringle Creek (NEON domain 11). This will allow for a comparative analysis of how biological source material and particle size affect the spatio-temporal distribution of DNA under fluctuating seasonal parameters. We hypothesize that fine-scale eDNA particles originating from labile tissues will exhibit increased transport distances and accelerated degradation rates relative to larger particles. Preliminary results from the DISTANCE project will advance mechanistic understanding of eDNA dynamics and inform the application of eDNA methodologies in lotic ecosystems nationwide.