Rewetting a dry soil has long been known to cause a burst of respiration (the “Birch Effect”). Hypothesized mechanisms for this involve: (1) release of cellular materials as a result of the rapid increase in water potential stress and (2) stimulating C-supply to microbes via physical processes. The balance of these factors is still not well understood, particularly in the contexts of multiple dry/wet cycles and of how resource and stress patterns vary through the soil profile. We evaluated the effects of multiple dry/wet cycles on surface and subsurface soils from a California annual grassland. Treatments included 4, 6, and 12 cycles that varied the length of the drying period between rewetting events. Respiration was monitored after each wetting event while extractable C and N, microbial biomass, and microbial activity were assayed initially, after the first rewetting event, and at the end of the experiment. Initially, microbial biomass and activity (respiration, dehydrogenase, and N mineralization) in subsurface soils were ca. 10% and 20% of surface soil levels. After multiple cycles, however, subsurface soil microbial biomass and activity were enhanced by up to 8-fold, even in comparison to the constantly moist treatment. By comparison, surface soil microbial biomass and activity were either moderately (i.e. 1.5 times increase) or not affected by wetting and drying. Drying and rewetting led to a cascade of responses (soluble C release, biomass growth, and enhanced activity) that mobilized and metabolized otherwise unavailable soil carbon, particularly in subsurface soils.