| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Jupiter |
A | DNS | Bulldog Invitational 2025 |
| 2 | Jupiter |
A | DNS | Bulldog Invitational 2025 |
| 3 | Dreyfoos |
A | DNS | St Andrews Varsity 3 |
| 4 | Jupiter Christian |
A | DNS | FHSAA 1A Region 4 |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Fort Pierce Central |
A | 3:36.21 | Cobra Relays 2025 |
| 2 | Sebastian River |
A | 3:34.48 | Jensen Beach Meet #2 |
| 3 | Atlantic Christian |
A | 3:27.18 | FHSAA State Track & Field Championships (1A) |
| 4 | Treasure Coast | A | 3:19.87 | FHSAA 4A Region 2 |
| 5 | Dwyer |
A | 3:24.28 | FHSAA 4A Region 2 |
| 6 | A | 3:30.14 | FHSAA 3A Region 4 | |
| 7 | Palm Beach Lakes |
A | 3:40.90 | Royal Palm Beach Invitational 2026 |
| 8 | Suncoast |
A | 3:36.23 | Royal Palm Beach Invitational 2025 |