The purpose of this study was to compare selected physical fitness and performance variables between National Collegiate Athletic Association (NCAA) Division I and II football players. The subjects included offensive and defensive starters, excluding kickers and punters from 26 NCAA Division I and 23 Division II teams. Offensive players were grouped and compared by the following positions:​ quarterback, running back, wide receiver, tight end, and line. Defensive players were grouped and compared by the following positions:​ line, linebackers, and backs. Division I players were better in 58 of 117 comparisons (p ≤ 0.01). Division II players were not found to be better in any of the variables studied.