We perform molecular dynamics simulations to investigate the effect of hydrogenation on the Poisson's ratio of graphene. It is found that the value of the Poisson's ratio of graphene can be effectively tuned from positive to negative by varying the percentage of hydrogenation. Specifically, the Poisson's ratio decreases with an increase in the percentage of hydrogenation, and reaches a minimum value of −0.04 when the percentage of hydrogenation is about 50%. The Poisson's ratio starts to increase upon a further increase of the percentage of hydrogenation. The appearance of a minimum negative Poisson's ratio in the hydrogenated graphene is attributed to the suppression of the hydrogenation-induced ripples during the stretching of graphene. Our results demonstrate that hydrogenation is a valuable approach for tuning the Poisson's ratio from positive to negative in graphene.