| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | East River |
B | 3:25.27 | Ocoee Relays |
| 2 | NSU University |
A | 3:25.15 | FHSAA State Track & Field Championships (2A/Para-Athlete) |
| 3 | Mainland |
A | 3:21.82 | FHSAA 3A Region 2 |
| 4 | Winter Park |
A | 3:21.08 | Metro East Championship 2025 |
| 5 | Mainland |
B | 3:21.82 | FHSAA 3A Region 2 |
| 6 | Wekiva |
A | 3:24.33 | Mustang Relays |
| 7 | East River |
A | 3:25.27 | Ocoee Relays |
| 8 | Flagler Palm Coast |
A | 3:26.44 |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Menendez |
A | 3:33.04 | George Janan Spruce Creek Invitational |
| 2 | Deltona |
A | 3:31.36 | Sanford-Seminole Coach McPhearson Invitational 2026 |
| 3 | West Port |
A | 3:30.00 | |
| 4 | West Port |
B | 3:28.00 | |
| 5 | Spruce Creek |
A | 3:29.09 | |
| 6 | Clay |
A | 3:30.96 | 2026 The Bulldog Classic Track and Field Meet |
| 7 | New Smyrna Beach |
A | 3:31.66 | Tocoi Creek Toro Showdown 2025 (Approved Qualifier) |
| 8 | University (OC) |
A | 3:42.26 | 5-Star Conference |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 3 | Orlando Christian Prep |
B | 3:59.46 | Hawk's Early Bird Invitational |
| 4 | Spruce Creek |
B | 3:45.09 | |
| 5 | Orlando Christian Prep |
A | 3:59.46 | Hawk's Early Bird Invitational |
| 6 | Menendez |
B | NT |