The 1965 Iowa State Cyclones football team represented Iowa State University in the Big Eight Conference during the 1965 NCAA University Division football season. In their eighth year under head coach Clay Stapleton, the Cyclones compiled a 5–4–1 record (3–3–1 against conference opponents), finished in fourth place in the conference, and outscored their opponents by a combined total of 178 to 147. They played their home games at Clyde Williams Field in Ames, Iowa. Steve Balkovec was the punter and placekicker.