Technical Abstract: Aerobic incubation methods have been widely used to assess soil nitrogen (N) mineralization, but standardized protocols are lacking. A silt loam soil (Catlin silt loam; fine-silty, mixed, superactive, mesic, Oxyaquic Arguidoll) was subjected to aerobic incubation at six USDA-ARS locations using a standardized protocol. Incubations were conducted at multiple temperatures, which were combined based on degree days (DD). Soil water was maintained at 60% water-filled pore space (WFPS; Constant) or allowed to fluctuate between 60 and 30% WFPS (Cycle). Soil subsamples were removed periodically and extracted in 2 M KCl; nitrate and ammonium concentrations in extracts were determined colorimetrically. For each location, the rate of soil organic matter N (SOMN) mineralization was estimated by regressing soil inorganic N (Ni) concentration on DD, using a linear (zero-order) model. When all data were included, the mineralization rate from four datasets was not statistically different, with a rate equivalent to 0.5 mg N/kg soil/ day. Soil incubated at two locations exhibited significantly higher SOMN mineralization rates. To assess whether this may have been due to pre-incubation conditions, time zero data were excluded and regression analysis was conducted again. Using this data subset, SOMN mineralization from five (of six) datasets was not significantly different. Fluctuating soil water reduced N mineralization rate at two (of four) locations by an average of 50%; fluctuating soil water content also substantially increased variability. This composite dataset demonstrates that standardization of aerobic incubation methodology is possible.