The genomic revolution has fundamentally changed how we survey biodiversity on earth. High-throughput sequencing (‘HTS’) platforms now enable the rapid sequencing of DNA from diverse kinds of environmental samples (termed ‘environmental DNA’ or ‘eDNA’). Coupling HTS with our ability to associate sequences from eDNA with a taxonomic name is called ‘eDNA metabarcoding’ and offers a powerful molecular tool capable of non-invasively surveying species richness from many ecosystems. Here, we review the use of eDNA metabarcoding for surveying animal and plant richness, and the challenges in using eDNA approaches to estimate relative abundance. We highlight eDNA applications in freshwater, marine, and terrestrial environments, and in this broad context, we distill what is known about the ability of different eDNA sample types to approximate richness in space and across time. We provide guiding questions for study design and discuss the eDNA metabarcoding workflow with a focus on primers and library preparation methods. We additionally discuss important criteria for consideration of bioinformatic filtering of data sets, with recommendations for increasing transparency. Finally, looking to the future, we discuss emerging applications of eDNA metabarcoding in ecology, conservation, invasion biology, biomonitor