Background/Aims: Excessive sedentary behaviour has been associated with childhood obesity.Schools offer an environment in which healthy lifestyle behaviours can be promoted and practised.Previous studies in older children (10-12 years) have shown that children are sedentary for around two-thirds of school-time.This study aims to investigate school effects on sedentary time in 5-6-year-old children, and the relationship between school-time allocation for breaks and physical education (PE) and sedentary time, within and outside of school hours.Methods: We used baseline data from 962 children from 50 schools in the West Midlands, England participating in the WAVES childhood obesity prevention trial.Sedentary time was measured using accelerometers worn for up to 5 days.School physical activity (PA) opportunities were identified through a school questionnaire.Adjusted random effects multilevel models were developed to determine associations between school PA opportunities and children's in-and out-of-school sedentary time.Intra-class correlation coefficients (ICCs) were calculated to estimate the proportion of variance attributed to between-school differences.Gender differences were explored.Results: On average children were sedentary for 30% of school-time (108.5 min/day), but total daily sedentary time was greater during weekend, compared to school days (464 vs 421 minutes).Significant variation was found between in-and out-of-school sedentary time that could be attributed to differences between schools.The strongest effect was for in-school-time (χ² = 72.50,p < 0.0001, ICC = 0.135) showing that 13.5% of total variation in school sedentary time is explained by school differences.Overall, there is a trend of decreasing school-time sedentary behaviour with increased school PA opportunities.However, gender-specific analyses suggest that school PA opportunities are somewhat associated with both less overall and out-of-school sedentary time in boys, but the opposite in girls.There was a significant positive association between school provision of PE and breaks and non-school sedentary time in girls (B = 8.89 min/day (1.00, 16.79), p = 0.027). Conclusions:The school PA environment is associated with children's sedentary time.The relationship between school PA opportunities and girls' sedentary behaviours is concerning and warrants further investigation.In addition, strategies to reduce children's sedentary time at weekends should be prioritised.