The aim of this study was to investigate the effect of post-activation potentiation (PAP), carbohydrate (CHO) mouth rinse, and the combination of both strategies on repeated sprint ability (RSA). Twenty male soccer players (age = 18.9 ± 0.9 years, body mass = 71.8 ± 5.2 kg, height = 178.2 ± 6.3 cm) randomly performed four experimental conditions before RSA test (six sets of 40 m): (I) placebo (PLA) control, (II) CHO mouth rinse (6% maltodextrin), (III) PAP + PLA, and (IV) PAP + CHO. The PAP protocol involved two sets of five repetitions (80% 1RM) of the back squat exercise. A one-way repeated measures analysis of variance followed by Bonferroni post hoc test was used to compare the experimental conditions. Results indicated that PAP + CHO and PAP + PLA had better results for the variables best sprint time, mean sprint time, and total sprint time compared with CHO and PLA (p<0.001; small effect size). No significant interaction between the experimental conditions was observed for the variable RSA performance decrement, and no significant difference between conditions, i.e., PAP + CHO vs. PAP + PLA and CHO vs. PLA control was found. In conclusion, PAP positively affects RSA performance in soccer players; however, the combination of PAP and