Soil carbon (C) and nitrogen (N) cycles are inextricably linked, yet the impacts of N availability upon soil C sequestration and turnover are poorly understood. According to stoichiometric theory, in the absence of nutrient limitation substrate decomposition will reach maximum rates, with C assimilated into microbial biomass at the expense of CO2 production. In this study, we added a 14C labelled low molecular weight substrate (glucose) to a sandy soil along with eleven increasing levels of N, phosphorus (P), and sulphur (S) in relative proportions as required for microbial biomass production. Adding a simple soluble substrate allowed us to explicitly examine changes in microbial transformations of added C, rather than changes resulting from extracellular enzyme activity or the extent of substrate decomposition. We hypothesized that as nutrient addition increased, an increasing proportion of the glucose-C provided would be incorporated into microbial biomass at the expense of CO2 production and stabilized as soil organic carbon (SOC). Instead, CO2 production from glucose-C increased significantly with nutrient addition without measurable changes in glucose-derived microbial biomass or SOC. This suggests that if there was greater glucose-derived microbial biomass produced under higher nutrient addition it was offset by a higher rate of microbial biomass turnover. We also found greater soil-derived microbial biomass at lower nutrient addition levels, potentially supporting the concept of microbial mining of soil organic matter (SOM) for nutrients under low nutrient availability. In conclusion, our data suggest that in a sandy soil with low capacity for physical protection of SOM, nutrient addition does not immediately promote C sequestration in the soil microbial community, and that the interaction between C stabilization and nutrient addition requires further work, especially for predicting ecosystem responses.