This study assessed the electrical activity of the masticatory muscles in both children with down syndrome (DS) and healthy children. After applying the inclusion and exclusion criteria, 30 patients aged between 7.9 and 11.8 years participated in the study. They were divided into two groups of 15: DS and non-DS. A DAB-Bluetooth device (Zebris Medical GmbH, Germany) was used to record the electromyographical (EMG) activity of the right and left temporal and of the right and left masseter muscles at rest and during maximum voluntary clenching (MVC). The asymmetry index between right and left masticatory muscle EMG activity was calculated for each position. The Mann–Whitney U test was applied to analyze the study results. There were no differences in the electrical activity of the temporal and masseter muscles at rest between the groups. During MVC, the asymmetry index for the masseter muscles was significantly higher in subjects with DS. The electrical potentials of the temporal and masseter muscles in children with DS were significantly lower compared to the corresponding parameters for healthy children when clenching.