| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 3 | Nittany Track and Field | B | Chan | Chan | Raish | Coulter | 2:50.00 |
| 4 | Nittany Track and Field | A | Croteau | Dunahay | Ulrich | Ranck | 2:50.00 |
| 5 | YMCA Of Indiana County | A | Herring | Gregory | Pruger | Neese | 3:02.04 |
| 6 | Nittany Track and Field | C | TAKAHASHI | Harpster | Edor | Feuer | 2:50.00 |