This study was performed using population-based data to analyze whether motor development in early life is different between singletons and twins in Japan. For better comparison and investigation, we divided the 2 groups into a group with a birthweight of 2.5 kg or greater (subgroup A) and a group with a birthweight of less than 2.5 kg (subgroup B), respectively. We analyzed the database of medical check-ups for children aged 3 years between April 2001 and July 2004. They received medical checkups at 4 months, 1.5, and 3 years of age. Children who were suspected of having neurological abnormality or disability were referred to specialists and excluded from the database. The data of 14,132 children were analyzed. Among these, 13,040 (92.3%) children were singletons in subgroup A, 75 (0.5%) were twins in subgroup A, 866 (6.1%) were singletons in subgroup B and 151 (1.1%) were twins in subgroup B. The mean age at achieving milestones was slower in twins of subgroup A for each developmental outcome than singletons of subgroup A, and the difference between twins and singletons was significant after adjustment for rolling over. On the contrary, after adjusting for a confounding factor (gestational age), singletons of subgroup B attained motor development facilitating walking independently slower than twins of subgroup B. There were different tendencies in the results regarding the motor development of subjects of subgroup A and that of subjects of subgroup B.