This study investigated how the ergonomic factors, such as hand dominance and postural selection, influenced on surgical performance regarding the changes of muscle activity. Twenty novices performed two virtual laparoscopic surgical training tasks and five target muscle activities were measured. Compared with using dominant hand, surgical skills performance using non-dominant hand increased muscle activities. Muscle fatigue is more likely induced in standing position than sitting position during practice. This study suggests an emerging need to focus on hand dominance during laparoscopic surgical training to address the impact of hand discrepancy on bimanual coordination. It is also important to pay attention on postural selection during training to reduce muscle fatigue, which possibly leads to injuries.