A simulation, optimization and analysis based on Witness software to study and analyze two-sided assembly line at the motorcycle assembly plant. The optimization process initiates with selection of company, product and data collection. The real environment based on company's initial scenario is modeled into Witness software and analyzed by using Cause-and-Effect diagram. Proposal of an improved solution such as manpower and layout planning with approach of Witness simulation is implemented on real case study in order to optimize performance of two-sided assembly line. Comparison are made between existing model and proposed model in order to evaluate the feasible efficiency of Witness application on the assembly line. The simulation results indicated that it minimized the occurrence of bottlenecks, reduced the assembly time and dramatically increased the final outputs. At the end of results, it successfully reduced the risk of getting low productivity and heighten the company's profits. It is envisage that the proposed model can be very useful for better productivity.