| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Penn State T&F Club | B | 43.50 | |
| 2 | Penn State T&F Club | A | 43.00 | |
| 3 | Lebanon Valley |
A | 41.90 | |
| 4 | Shippensburg |
A | 40.89 | 2025 Colonial Relays |
| 5 | Cortland St. |
A | 41.15 | Cortland Classic |
| 6 | York (Pa.) |
A | 42.20 | |
| 7 | PSU-Harrisburg |
A | 43.11 | 43.11 Mason-Dixon Track %26 Field Invitational |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 3 | Messiah |
A | 46.90 | 2025 Messiah Multi/Invitational |
| 4 | York (Pa.) |
B | 43.70 | |
| 5 | Albright |
A | 45.46 | Purple & Gold Open |