The aim of the study was to quantify the activity as recorded by electromyography during ergometer cycling in eleven different muscles of the lower extremity. Eleven healthy subjects rode in twelve different ways at different work-load, pedalling rate, saddle height and pedal foot position. Vastus medialis and lateralis, gastrocnemius medialis and lateralis and the soleus muscle were the most activated muscles. Changes in muscle activity during different calibrations were studied in eight of the eleven muscles. An increase in work-load significantly increased the mean maximum activity in all the eight muscles investigated. An increase of the pedalling rate increased the activity in the gluteus maximus, gluteus medius, vastus medialis, medial hamstring, gastrocnemius medialis and soleus muscles. An increase of the saddle height increased the muscle activity in the gluteus medius, medial hamstring and gastrocnemius medialis muscles. Use of a posterior pedal foot position increased the activity in the gluteus medius and rectus femoris muscles, and decreased the activity in the soleus muscle.