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| 07.10.2009 | 19:30 | CD Lagun Onak | - | AD Alcorcón | 1:2 | |
| 20:45 | UD Salamanca | - | FC Cartagena | 2:0 | ||
| 20:45 | Unión Deportiva Puertollano | - | UE Sant Andreu | 2:1 | ||
| 21:00 | Córdoba CF | - | Rayo Vallecano | 0:1 ap | ||
| 21:00 | Recreativo Huelva | - | UD Las Palmas | 2:1 | ||
| 21:00 | Hércules CF | - | SD Huesca | 2:0 | ||
| 21:00 | Girona FC | - | RC Celta | 1:3 | ||
| 21:00 | Mérida UD | - | CD Alcoyano | 1:3 | ||
| 21:00 | CF Atlético Ciudad | - | Polideportivo Ejido | 5:4 tab | ||
| 21:00 | UD Marbella | - | Alicante CF | 2:1 ap | ||
| 21:00 | Ontinyent CF | - | Cultural Leonesa | 3:5 tab |
