BioNetFit can be used on a stand-alone computer or a Linux-based cluster. It is compatible with BioNetGen and NFsim. Thus, the various simulation capabilities provided by these software packages (generate-first deterministic, stochastic and hybrid methods and network-free stochastic methods) can be used with BioNetFit, which implements a genetic algorithm. BioNetFit can perform a global fit, meaning that it can consider multiple datasets in the course of a fitting run. Preprocessing of data (for example, normalization of the average of the measurement values in each dataset) is advised when considering multiple datasets. BioNetFit can consider steady-state dose-response data and/or time-series data.