ABSTRACT: Children's after-school physical activity (PA) and sedentary behaviours (SB) are not well understood, despite the potential this period holds for intervention. This study aimed to describe children's after-school physical activity and sedentary behaviours; establish the contribution this makes to daily participation and to achieving physical activity and sedentary behaviours guidelines; and to determine the association between after-school moderate- to vigorous-intensity physical activity (MVPA), screen-based sedentary behaviours and achieving the physical activity and sedentary behaviour guidelines.Children (n = 406, mean age 8.1 years, 58% girls) wore an ActiGraph GT3X accelerometer. The percentage of time and minutes spent sedentary (SED), in light- physical activity (LPA) and MVPA between the end-of-school and 6pm (weekdays) was calculated. Parents (n = 318, 40 years, 89% female) proxy-reported their child's after-school participation in screen-based sedentary behaviours. The contribution that after-school SED, LPA, MVPA, and screen-based sedentary behaviours made to daily levels, and that after-school MVPA and screen-based sedentary behaviours made to achieving the physical activity/sedentary behaviour guidelines was calculated. Regression analysis determined the association between after-school MVPA and screen-based sedentary behaviours and achieving the physical activity/sedentary behaviours guidelines.Children spent 54% of the after-school period SED, and this accounted for 21% of children's daily SED levels. Boys spent a greater percentage of time in MVPA than girls (14.9% vs. 13.6%; p<0.05), but this made a smaller contribution to their daily levels (27.6% vs 29.8%; p<0.05). After school, boys and girls respectively performed 18.8 minutes and 16.7 minutes of MVPA, which is 31.4% and 27.8% of the MVPA (p<0.05) required to achieve the physical activity guidelines. Children spent 96 minutes in screen-based sedentary behaviours, contributing to 84% of their daily screen-based sedentary behaviours and 80% of the sedentary behaviour guidelines. After-school MVPA was positively associated with achieving the physical activity guidelines (OR: 1.31, 95%CI 1.18, 1.44, p<0.05), and after-school screen-based sedentary behaviours were negatively associated with achieving the sedentary behaviours guidelines (OR: 0.97, 95%CI: 0.96, 0.97, p<0.05).The after-school period plays a critical role in the accumulation of children's physical activity and sedentary behaviours. Small changes to after-school behaviours can have large impacts on children's daily behaviours levels and likelihood of meeting the recommended levels of physical activity and sedentary behaviour. Therefore interventions should target reducing after-school sedentary behaviours and increasing physical activity.