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Biological nitrogen fixation converts inert di-nitrogen gas into bioavailable nitrogen and can be an important source of bioavailable nitrogen to organisms. This dataset synthesizes the aquatic nitrogen fixation rate measurements across inland and coastal waters. Data were derived from papers and datasets published by April 2022 and include rates measured using the acetylene reduction assay (ARA), 15N2 labeling, or the N2/Ar technique. The dataset is comprised of 4793 nitrogen fixation rates measurements from 267 studies, and is structured into four tables: 1) a reference table with sources from which data were extracted, 2) a rates table with nitrogen fixation rates that includes habitat, substrate, geographic coordinates, and method of measuring N2 fixation rates, 3) a table with supporting environmental and chemical data for a subset of the rate measurements when data were available, and 4) a data dictionary with definitions for each variable in each data table. This dataset was compiled and curated by the NSF-funded Aquatic Nitrogen Fixation Research Coordination Network (award number 2015825).