This dataset contains the results of a five-day field excursion which the extent to which eDNA sampling can capture the diversity of a region with highly heterogeneous habitat patches across a wide elevation gradient through multiple hydrological catchments of the Swiss Alps. Using peristaltic pumps, we filtered 60 L of water at five sites per catchment for a total volume of 1 800 L. Using an eDNA metabarcoding approach focusing on vertebrates and plants, we detected 86 vertebrate taxa spanning 41 families and 263 plant taxa spanning 79 families across ten catchments. This dataset includes two sets of data. The first (Genomic data) includes all the necessary data for the bioinformatic pipeline, whereas the second (Analysis Figures) contains tidied data and scripts for the reproduction of all figures/analyses in the article describing this study.