| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Albright |
A | 45.46 | Purple & Gold Open |
| 2 | DeSales |
A | 43.68 | DeSales Quad Meet |
| 3 | Widener |
A | 42.14 | 2025 Paul Donahue Invitational at Widener University |
| 4 | A | 44.01 | 2025 Kings College Monarch Open | |
| 5 | Eastern |
A | 42.23 | Danny Curran Invite |
| 6 | A | 42.53 | Penn Relays | |
| 7 | A | 42.36 | 2025 Kings College Monarch Open | |
| 8 | Messiah |
A | 45.64 | 20th Annual Paul Kaiser Classic |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Delaware Valley |
A | 42.79 | Danny Curran Invite |
| 2 | Stevens |
A | 42.61 | 2025 Coach P Invite |
| 3 | Misericordia |
A | 42.26 | Penn Relays |
| 4 | York (Pa.) |
A | 42.29 | 20th Annual Paul Kaiser Classic |
| 5 | Lebanon Valley |
A | 42.00 | 2025 Messiah Multi/Invitational |
| 6 | Alvernia |
A | 44.43 | York College Twilight |
| 7 | Stevenson |
A | 44.11 | Towson Invitational |