This paper presents a new robot calibration algorithm with joint stiffness parameters for the enhanced positioning accuracy of industrial robot manipulators. This work is towards on-going development of an industrial robot calibration software which is able to identify both the kinematic and non-kinematic robot parameters. In this paper, the conventional kinematic calibration and its important considerations are briefly described first. Then, a new robot calibration algorithm which simultaneously identifies both the kinematic and joint stiffness parameters is presented and explained through a computer simulation with a 2 DOF manipulator. Finally, the developed algorithm is implemented to Hyundai HX165 robot and its resulting improvement of the positioning accuracy is addressed.