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Biological nitrogen fixation is the conversion of dinitrogen (N2) gas into bioavailable nitrogen by microorganisms with consequences for primary production, ecosystem function, and global climate. Here we present a compiled dataset of 4793 nitrogen fixation (N2-fixation) rates measured in the water column and benthos of inland and coastal systems via the acetylene reduction assay, 15N2 labeling, or N2/Ar technique. While the data are distributed across seven continents, most observations (88%) are from the northern hemisphere. 15N2 labeling accounted for 67% of water column measurements, while the acetylene reduction assay accounted for 81% of benthic N2-fixation observations. Dataset median area-, volume-, and mass-normalized N2-fixation rates are 7.1 μmol N2-N m−2 h−1, 2.3 × 10−4 μmol N2-N L−1 h−1, and 4.8 × 10−4 μmol N2-N g−1 h−1, respectively. This dataset will facilitate future efforts to study and scale N2-fixation contributions across inland and coastal aquatic environments.