Aims: The aim of this study was to reveal the aerobic and facultative anaerobic bacterial diversity in the supragingival biofilms of healthy caries-free children.Methodology and results: Biofilm sampling was performed by a specialist in the morning at least 12 h after tooth brushing from 6-10 years old children who came to Suleyman Demirel University Pedodontics Clinic. Samples were cultured on selected media. Purified isolates were identified according to 16S rRNA gene sequences. Totally 180 bacterial isolates had obtained. These isolates were identified as 36 different bacterial species belonging to Firmicutes, Proteobacteria and Actinobacteria. Species with the highest number of isolates were Lactobacillus and Staphylococcus. Conclusion, significance and impact of study: In this study, a total of 180 bacterial isolates had obtained. These isolates were identified as 36 different bacterial species belonging to 3 bacterial phyla (Firmicutes, Proteobacteria and Actinobacteria) and 9 bacterial genera. Although our study has certain limitations because it is dependent on culture, since there is no study previously conducted in Turkey to our knowledge, it contains important findings. The research findings reported in this paper will serve as the foundation for studies to be conducted to understand the oral colonization and biofilm dynamics of children and to take protective measures accordingly.