Semi-sealed preservation of soil samples at different moisture of 4% and 23%, respectively, was simulated to observe the variations of soil microbial communities and determine the contents and isotopic compositions of the total organic carbon and total nitrogen on the 7th and 30th day, respectively. The results show that during preservation, the quantity of microbial communities tended to increase first and then decrease, with a wider variation range at higher moisture (23%). At the moisture content of 23%, the microbial communities became more active on the 7th day, but less after 30 days, and their activity was stable with little fluctuation at the moisture content of 4%. However, there were no significant changes in the contents and isotopic compositions of the total organic carbon and total nitrogen. During preservation, the responses of soil microbes to the environment are more sensitive to changes in the total nitrogen and organic carbon contents. It is thus suggested that the variations of microbial communities have not exerted remarkable impacts on the isotope compositions of the total nitrogen and total organic carbon.