In the first game of his sophomore campaign, Barkley rushed for 105 yards and a touchdown in a 33–13 win over Kent State. In week two, in a renewed rivalry game against Pitt, Penn State trailed at one point in the second quarter 28–7. Barkley scored five touchdowns, his fifth coming with 5:00 left in the fourth quarter, to bring the Nittany Lions within three points. However, quarterback Trace McSorley threw an interception in the Pitt end zone with 1:41 left, that denied the Nittany Lions' hopes of completing the comeback. In week five against Minnesota, in what would become a recurring theme for the 2016 Penn State football team, Penn State had to come back from another halftime deficit. In overtime, on Penn State's first play, Barkley ended the game with a 25-yard touchdown run. Before that carry, Minnesota had held him to only 38 yards on 19 carries. In week six against Maryland, Barkley had his first ever 200-yard rushing game in college. He finished with 31 carries for 202 yards and one touchdown. In week seven, in a matchup versus Ohio State, Barkley rushed for 99 yards on 12 carries with a long of 37 in a 24–21 win over the no. 2 Buckeyes. In week eight, Barkley and no. 24 Penn State scored a season high 62 points versus Purdue. Barkley achieved a career-high 207 rushing yards with a career-high long of 81 yards and two touchdowns, along with 70 yards of receiving, giving him a total of 277 yards on the day. Penn State defeated Purdue 62–24 and improved to 6–2 and 4–1 in Big Ten play.