The stability of biochar in soil is of importance if it is to be used for carbon sequestration and long-term improvement of soil properties. It is well known that a significant fraction of biochar is highly stable in soil, but carbon dioxide (CO2) is also released immediately after application. This study investigated the nature of the early release of CO2 and the degree to which stabilizing mechanisms protect biochar from microbial attack. Incubations of 14C-labelled biochar produced at different temperatures were performed in soils with different clay contents and in sterilized and non-sterilized soils. It emerged that carbonate may be concentrated or form during or after biochar production, resulting in significant carbonate contents. If CO2 released from carbonates in short-term experiments is misinterpreted as mineralization of biochar, the impact of this process may be significantly over-estimated. In addition to the CO2 released from carbonates, there appears to be a labile fraction of biochar that is oxidized quickly during the first days of incubation, probably by both abiotic and biotic processes. Later in the incubation, biotic mineralization appears to be the primary cause of CO2 evolution. Finally, we found that both production temperature and clay content affect biochar mineralization. As protective mechanisms hypothesized to prevent degradation of organic matter in soil usually implicate clay, we conclude that biochar is likely to be protected from mineralization during the early stages of incubation by its own recalcitrant chemical and physical nature as well as by physical protective mechanisms.