Abstract: Priming effect is one of the important mechanisms regulating soil organic matter decomposition. However, the variation of priming effects in different soil layers remains unclear. In this study, we conducted a 30-day incubation experiment using no-tillage black soil from northeastern China. ¹³C-glucose and dynamic CO₂ trapping methods were employed to investigate soil organic carbon (SOC) mineralization rates and the priming effect of the added ¹³C-glucose in the upper soil layer (0-10 cm) and the lower soil layer (30-40 cm). Our results showed that the cumulative SOC-specific mineralization rate in the upper layer was similar to that in the lower layer soil without glucose addition. Glucose addition significantly altered the mineralization rates in both layers, resulting in a positive priming effect (36.7%) in the upper layer but a negative priming effect (-12.4%) in the lower layer. The cumulative priming effect during the 30-day incubation was 3.24 mg C·g^-1 SOC for the upper layer soil and -1.24 mg C·g^-1 SOC for the lower layer soil. There was still a net SOC increase, even with positive priming effects in the upper layer soil. This was due to considerable amount of added glucose-C remained un-mineralized in the soil which would compensate the carbon loss from priming effects. Overall, our results demonstrated that the magnitude and direction of priming effects might differ between soil layers. Our findings contribute to a better understanding of the effects of conservation tillage practices (no-tillage and straw incorporation) on soil organic matter dynamics in agroecosystems.