The rotation matrix estimation problem is a keypoint for mobile robot localization, navigation, and control. Based on the quaternion theory and the epipolar geometry, an extended Kalman filter (EKF) algorithm is proposed to estimate the rotation matrix by using a single-axis gyroscope and the image points correspondence from a monocular camera. The experimental results show that the precision of mobile robot s yaw angle estimated by the proposed EKF algorithm is much better than the results given by the image-only and gyroscope-only method, which demonstrates that our method is a preferable way to estimate the rotation for the autonomous mobile robot applications.