The purpose of this study was to examine the difference in children\'s spatial ability between experimental group used educational software and comparative group used mathematical play material for spatial concept. The subjects of this study were 50 five-years old children and Spatial Ability Test was administered to measure spatial ability of subjects, which was composed of five sub types of spatial ability including eye-motor coordination, figure-ground perception, inference on space, space relationship, and visual memorization and recollection. This study found statistically significant differences in spatial ability between experimental group and comparative group. Children who played with educational software scored higher in spatial ability test than comparative group who played with mathematical play material. There was the statistically significant difference between experimental group and comparative group in figure-ground perception and visual memorization and recollection among sub types of spatial ability. Moreover, there was no statistically significant difference in spatial ability between boys and girls but boys who played with educational software scored higher in spatial ability compared to scores of boys in comparative group.