Nutrient uptake length is an important parameter for quantifying nutrient cycling in streams. Although nutrient tracer additions are the preferred method for measuring uptake length under ambient nutrient concentrations, short-term nutrient addition experiments have more frequently been used to estimate uptake length in streams. Theoretical analysis of the relationship between uptake length determined by nutrient addition experiments (S-w') and uptake length determined by tracer additions (S-w) predicted that S-w' should be consistently longer than S-w, and that the overestimate of uptake length by S-w' should be related to the level of nutrient addition above ambient concentrations and the degree of nutrient limitation. To test these predictions, we used data from an interbiome study of NH4+ uptake length in which (NH4+)-N-15 tracer and short-term NH4+ addition experiments were performed in 10 streams using a uniform experimental approach. The experimental results largely confirmed the theoretical predictions: S-W' was consistently longer than S,a, and S,,':S,, ratios were directly related to the level of NH4+ addition and to indicators of N limitation. The experimentally derived S-w':S-w ratios were used with the theoretical results to infer the N limitation status of each stream. Together, the theoretical and experimental results showed that tracer experiments should be used whenever possible to determine nutrient uptake length in streams. Nutrient addition experiments may be useful for comparing uptake lengths between different streams or different times in the same stream, however, provided that nutrient additions are kept as low as possible and of similar magnitude.