A seepage investigation was conducted by the U.S. Geological Survey, in cooperation with the New Mexico Interstate Stream Commission, along an approximately 106-mile reach of the Rio Grande from below Caballo Reservoir, New Mexico, to El Paso, Texas, during June 26–28, 2012, to determine gain or loss of streamflow due to seepage to or from the river channel. Discharge measurements were made during the irrigation season at high flow including 5 sites along the Rio Grande, 5 diversions, and 63 inflows. The net gain or loss of flow in the river channel was computed for four reaches within the 106-mile reach of the Rio Grande. The normalized percentage difference was computed for each reach to determine the difference between discharge measured at upstream and downstream sites, and the normalized percentage uncertainty was computed to determine if a computed gain or loss exceeded cumulative uncertainty associated with measurement of discharge.