In this paper, the theoretical analysis and simulating calculation were conducted for a basic two-stage semiconductor thermoelectric module, which contains one thermocouple in the second stage and several thermocouples in the first stage. The study focused on the configuration of the two-stage semiconductor thermoelectric cooler, especially investigating the influences of some parameters, such as the current I1 of the first stage, the area A1 of every thermocouple and the number n of thermocouples in the first stage, on the cooling performance of the module. The obtained results of analysis indicate that changing the current I1 of the first stage, the area A1 of thermocouples and the number n of thermocouples in the first stage can improve the cooling performance of the module. These results can be used to optimize the configuration of the two-stage semiconductor thermoelectric module and provide guides for the design and application of thermoelectric cooler.