The early routing of the Midland Trail, from east to west, began in either New York City or Washington, D.C., and continued through Richmond and Clifton Forge, Virginia to Charleston, West Virginia, and passed on through Morehead, Kentucky, to Lexington, Kentucky; Louisville, Kentucky; Vincennes, Indiana; Salem, Illinois; St. Louis, Missouri; Sedalia, Missouri; Kansas City, Missouri, and Topeka, Kansas; to Limon, Colorado, and then on to Denver, Colorado.
Past Salt Lake City, the routing moved southward across the Salt Lake Desert on the same routing as the Lincoln Highway through Iosepa, Utah, Orr's Ranch, Fish Springs Ranch, and Ibapah, Utah.
In central Nevada, the highway continued across the Great Basin Desert through Ely and Tonopah then turning south at Goldfield in the Amargosa Desert and then west into California at Lida and over the Inyo Mountains and White Mountains through Westgard Pass.
At the junction in Big Pine, California in the Owens Valley, the original routing then split into four options: Following a major realignment of the route and assumption into the state highway system around 1922, the main Midland Trail alignment in California bypassed early stagecoach-era stops at Freeman and Willow Springs and at the Neuralia railroad siding, and now routed through Red Rock Canyon to Mojave.
Various alignments of this portion of the trail followed the late 19th century Twenty-mule team roads built to haul gold from the Cerro Gordo Mines across the Mojave Desert.