The purpose of this study was to investigate the effects of increased saddle height on the length and activity pattern of vastus lateralis (VL) and biceps femoris (BF) muscle. To compare the effects of increased saddle height, Preferred (self-selected height of subject) and High saddle height (approximately 5 % higher saddle height than self-selected) were used. Seven elite cyclists (career: 16.1±8.5 years) participated in 3 min. sub-maximal pedaling tests under the same cadence (90 RPM) and pedaling power (150 W). Hip and knee joint angles, and the length and activity of VL and BF were compared by measuring 3D motion and electromyography (EMG) data. Results showed that there were significant differences in peak extension timing of the hip joint angle and the range of motion of the hip and knee join between different saddle heights. Although there were significant differences in muscle length of both muscles with increasing saddle height, the timing and amount of muscle activity differed only at the CF. These findings suggest that the timing and amount of bi-articular muscle activity (i.e. BF) can be altered by changing the saddle height. For practically applying these results, further study is necessary to evaluate the effects of various cadence and the pedaling power with various saddle heights.