Robot positioning accuracy plays an important role in industrial automation applications. In this paper, a method is proposed for the improvement of robot accuracy with an optical tracking system that integrates a least-square numerical algorithm for the identification of kinematic parameters. In the process of establishing the system kinematics model, the positioning errors of the tool and the robot base, and the errors of the Denavit-Hartenberg parameters are all considered. In addition, the linear dependence among the parameters is analyzed. Numerical simulation based on a 6-axis UR robot is performed to validate the effectiveness of the proposed method. Then, the method is implemented on the actual robot, and the experimental results show that the robots can reach desired poses with an accuracy of ±0.35 mm for position and ±0.07° for orientation. Benefitting from the optical tracking system, the proposed procedure can be easily automated to improve the robot accuracy for applications requiring high positioning accuracy such as riveting, drill, and precise assembly.