This research was conducted to analyze play and playmate preferences of preschool girls and boys during free play time. The study group consisted of 48-66 months old preschool children. There were ten girls, seven boys and a preschool teacher in the study group. They were all selected from a public preschool in Cukurova, a district of Adana a city in the south of Turkey in the fall semester of 2014 to 2015 academic year. For two weeks children were observed during their free play time. In addition, the children and teacher were interviewed to determine the play and playmate preferences of the children. An interview with each child took approximately 5 to 6 minutes, whereas the interview with the teacher took 20 minutes. In this research, content analysis method was applied at the data analysis phase. The outcomes of the analysis indicated that there were differences between play area, type of play and playmate preferences between girls and boys. Research indicated that girls usually opt for dramatic (pretend) play such as playing house, role play and gets in role as doctor, mother or kid; while boys prefer to play with blocks and lego for construction and building purposes (block center). The research results show gender effects on girls and boys play behaviors.