It is of great importance to understand how terrestrial ecosystems will respond to global changes. However, most experimental approaches have focused on single factors. In natural systems, moisture and temperature often change simultaneously, and they can interact and shape microbial responses. Even though soil moisture and temperature are very important factors controlling microbial activity, there is disagreement on the dependence of microbial rates on temperature and moisture as well as their sensitivity when both variables change simultaneously. Here we created a moisture gradient and determined high resolution intrinsic temperature dependences for bacterial and fungal growth rates as well as respiration rates. We found that microbial rates decreased with lower moisture and increased with higher temperatures until optimum values. Additionally, we found independence between temperature and moisture as rate modifiers. We also found that temperature sensitivities (Q10) for microbial growth and respiration were not affected by changes in moisture. This provided an experimental framework to validate assumptions of temperature and moisture rate modifiers used in ecosystem and global cycling models (GCMs).