Environmental DNA (eDNA) is a rapidly emerging data source with the potential to support environmental monitoring and biodiversity conservation around the world. Current efforts to standardize eDNA methods and reporting are aimed at strengthening credibility and supporting adoption. In doing this, however, researchers must be mindful of diverse capacities and ecological contexts both regionally and around the world. The objective of our research is to understand how international standards for eDNA may support or hinder the uptake of eDNA methods and tools for conservation and biodiversity work. This was accomplished through two interactive workshops that brought together eDNA researchers and practitioners from around the world to surface broad and specific barriers to uptake of eDNA methods and tools.