The purpose of this study was to determine the effect of gender on changes in electromyographic (EMG) signal characteristics of the quadriceps muscles with increasing force and with fatigue. A total of fourteen healthy adults (seven men, seven women) participated in the study. Subjects had to perform isometric ramp contractions in knee extension with the force gradually increasing from 0 to 100% of the maximal voluntary contraction (MVC) in a 6-s period. Subjects then performed a fatigue task, consisting of a sustained maximum isometric knee extension contraction held until force decreased below 50% of the pre-fatigue MVC. Subjects also performed a single ramp contraction immediately after the fatigue task. The Root Mean Square (RMS) amplitude, mean power frequency (MPF) and median frequency (MF) of EMG signals obtained from the vastus lateralis, vastus medialis and rectus femoris were calculated at nine different force levels from the ramp contractions (10, 20, 30, 40, 50, 60, 70, 80 and 90% MVC), as well as every 5 s during the fatigue task. The main results were a more pronounced increase in EMG RMS amplitude for the three muscles and in MPF for the VL muscle with force in men compared with women. No significant effect of gender was found with regards to fatigue. These observations most likely reflect a moderately greater type II fiber content and/or area in the VL muscle of men compared to that of women.