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The aim of this cadaveric study was to evaluate the intraoperative kinematics of the native knee including two-dimensional translation of the femur using a navigation system. Eight native knees of 4 fresh-frozen whole-body cadavers were used for the study. The kinematics of each knee were analyzed intraoperatively using the navigation system. Although anterior-posterior translation could not be assessed directly, it could be calculated using a formula derived from the parameters in the navigation system. The native knee showed external rotation of the femur in early knee flexion, transient internal rotation in mid flexion, and gradual external rotation in late flexion. There was no marked change in the coronal rotation angle of the mechanical axis during knee flexion. The femoral center moved anteriorly in early knee flexion and posteriorly in late flexion. The distance moved in the medial-lateral direction was relatively smaller than that in the anterior-posterior direction. Two-dimensional translation of the surgical epicondylar axis showed a medial pivot-like motion. In this cadaveric study, the kinematics of the native knee, including two-dimensional translation of the femur, could be satisfactorily assessed intraoperatively using a navigation system. The intraoperative kinematics of the knee can be analyzed in more detail using this methodology.