In this study the problem formulation and solution technique using genetic algorithms for optimization design of laminate composite bicycle frame, with respect to its stacking sequence, was introduced. Tsai-Wu Criteria was applied to be as objective functions. The stacking sequence optimization, which deals with the discrete ply-angle, was used for development of composite bicycle frame. For optimization, each frame was divided into six parts; head tube, seat tube, top tube, bottom tube, seat stay, and chain stay. As they produces different stress distribution pattern among the parts, the unique stacking sequence should be applied for each part, and optimized with finite element method in two different loading conditions, level loading and vertical loading. Then the most effective stacking sequence of composite frame was decided, having the lowest failure index. Numerical results showed that the optimal stacking sequence is found with fewer evaluations of objective function than expected with size of feasible region, which shows the algorithm can be effectively used for lay-up optimization of composite bicycle.