Abstract Objective To evaluate asymmetry in placement of bilateral skin markers on horses and to determine effect of asymmetric skin marker placement on kinematic variables for trotting horses. Animals 10 horses for evaluation of asymmetry in marker placement; 1 horse for evaluation of effects on kinematic variables. Procedure Asymmetry in marker placement was assessed by attaching markers to horses and comparing radiographs of left and right limbs. An experimental model was developed to determine effects on kinematic variables; accuracy of the model was validated experimentally. Using kinematic data from a clinically normal trotting horse as reference data, effects of asymmetric marker placement on vertical displacement-time and joint angle-time diagrams were determined by use of the model. Results Asymmetry of placement was < 1 cm for markers on the distal portions of the limbs and < 2 cm for markers on the proximal portions. Asymmetric marker placement did not alter general shapes of the vertical displacement-time and joint angle-time curves. In most instances, largest differences in vertical displacement attributable to asymmetric marker placement were equal to or less than magnitude of the asymmetry of placement. Alterations in joint angle-time curves were mainly a result of shifting of the curves on the Y axis. Joint range of motion was only slightly changed by asymmetric marker placement, but maximum flexion and extension angles were greatly altered. Conclusions and Clinical Relevance Some kinematic variables can be greatly altered by small differences in skin marker placement. Such effects should be taken into account when evaluating kinematic data for sound and lame horses. (Am J Vet Res 1998;59:938–944)