Work-related musculoskeletal disorders are increasing due to industrialisation of work processes. Task variation has been suggested as potential intervention. The objectives of this study were to investigate, first, the influence of task variation on electromyographic (EMG) manifestations of shoulder muscle fatigue and discomfort; second, noticeable postural shoulder changes over time; third, if the association between task variation and EMG might be biased by postural changes. Outcome parameters were recorded using multichannel EMG, Optotrak and the Borg scale. Fourteen participants performed a one-hour repetitive Pegboard task in one continuous and two interrupted conditions with rest and a pick-and-place task, respectively. Manifestations of shoulder muscle fatigue and discomfort feelings were observed throughout the conditions but these were not significantly influenced by task variation. After correction for joint angles, the relation between task variation and EMG was significantly biased but significant effects of task variation remained absent. Practitioner Summary: Comparing a one-hour continuous, repetitive Pegboard task with two interrupted conditions revealed no significant influences of task variation. We did observe that the relation between task variation and EMG was biased by posture and therefore advise taking account for posture when investigating manifestations of muscle fatigue in assembly tasks.