Abstract. Temperature and moisture are primary environmental drivers of soil organic matter (SOM) decomposition, and the development of a better understanding fo their roles in this process through depth in soils is needed. The objective of this research is to independently assess the roles of temperature and moisture in driving heterotrophic soil respiration for shallow and deep soils in a temperate red spruce forest. Minimally disturbed soil cores from shallow (0–25 cm) and deep (25–50 cm) layers were extracted from a 20 yr old red spruce stand and were then transferred to a climate chamber where they were incubated for 3 months under constant and diurnal temperature regimes. Soils were subjected to different watering treatments representing a full range of water contents. Temperature, moisture, and CO₂ surface flux were assessed daily for all soils and continuously on a subset of the microcosms. The results from this study indicate that shallow soils dominate the contribution to surface flux (90%) and respond more predictably to moisture than deep soils. An optimum moisture range of 0.15 to 0.60 water-filled pore space was observed for microbial SOM decomposition in shallow cores across which a relatively invariant temperature sensitivity was observed. For soil moisture conditions experienced by most field sites in this region, flux-temperature relationships alone can be used to reasonably estimate heterotrophic respiration, as in this range moisture does not alter flux, with the exception of rewetting events along the lower part of this optimal range. Outside this range, however, soil moisture determines SOM decomposition rates.