"Mad" Madeleine Mayflower is a professional biker that, after competing for more than twenty years, decides to take a break from the race circuit, traveling with her motorcycle to explore the historical route 66 alongside her grandfather.

Route 66 is a straightforward hidden object game, where the objective is to find all the items from a list at the bottom of the screen, on picturesque locations around the United States filled with assorted paraphernalia. The hint button is a badge at the bottom-right, that shows the position of one of the required objects when activated.

Three mini-games appear between each location, giving extra hints to the player when completed. They are implementations of classic games like Simon with checkers pieces, a matching game with bottle taps and Reversi/Othello.