Fly Sampling Protocol

In recent years, invertebrate-derived DNA (iDNA) has emerged as a powerful tool for non-invasive biodiversity monitoring. This protocol outlines a standardized method for the collection of flies to assess local vertebrate diversity through environmental DNA (eDNA) analysis. By exploiting the natural behavior of carrion- and dung-feeding flies, which interact closely with vertebrate tissues and excreta, we obtain vertebrate DNA traces for downstream molecular analysis. The protocol includes guidelines for trap placement, bait selection, fly handling, and preservation methods. This approach enables efficient, scalable, and low-impact monitoring of vertebrate communities, with particular relevance for remote or logistically challenging ecosystems. The use of fly-derived eDNA represents a promising addition to the biodiversity assessment toolbox, complementing traditional survey methods and enhancing the detection of elusive or rare species.