The Environmental DNA Protocol Development Guide for Biomonitoring and Environmental DNA test validation guidelines were developed as a collaborative project, funded by the Australian Government Department of Agriculture, Fisheries and Forestry, and led by Dr Alejandro Trujillo-Gonzalez of the Australian National eDNA Reference Centre at the University of Canberra and Dr Maarten De Brauwer of CSIRO. More than 50 experts from across Australia and New Zealand were involved in the development of these guidelines and provided feedback during various stages of the process. These guidelines harmonise minimum quality standards to develop eDNA-based protocols and tests for biomonitoring applications. Both documents will be reviewed yearly to improve accuracy and clarity with current research and innovation.