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| H | Recreativo Huelva | - | Sporting Gijón | 1:1 (1:0) | ||
| R | Sporting Gijón | - | Recreativo Huelva | 2:4 (1:1, 1:1, 1:1) i.E. | ||
| H: 27.10.2009 20:00 | R: 10.11.2009 20:00 | Sieger: Recreativo Huelva | ||||
| H | CF Atlético Ciudad | - | Sevilla FC | 2:4 (2:4) | ||
| R | Sevilla FC | - | CF Atlético Ciudad | 5:1 (0:0) | ||
| H: 27.10.2009 20:00 | R: 10.11.2009 20:00 | Sieger: Sevilla FC | ||||
| H | UD Marbella | - | Atlético Madrid | 0:2 (0:1) | ||
| R | Atlético Madrid | - | UD Marbella | 6:0 (4:0) | ||
| H: 27.10.2009 20:00 | R: 10.11.2009 19:00 | Sieger: Atlético Madrid | ||||
| H | AD Alcorcón | - | Real Madrid | 4:0 (3:0) | ||
| R | Real Madrid | - | AD Alcorcón | 1:0 (0:0) | ||
| H: 27.10.2009 22:00 | R: 10.11.2009 20:00 | Sieger: AD Alcorcón | ||||
| H | Real Zaragoza | - | Málaga CF | 1:1 (1:0) | ||
| R | Málaga CF | - | Real Zaragoza | 0:0 (0:0) | ||
| H: 28.10.2009 20:00 | R: 10.11.2009 20:00 | Sieger: Málaga CF | ||||
| H | Xerez CD | - | CA Osasuna | 1:2 (0:2) | ||
| R | CA Osasuna | - | Xerez CD | 1:0 (0:0) | ||
| H: 28.10.2009 20:00 | R: 10.11.2009 20:00 | Sieger: CA Osasuna | ||||
| H | Rayo Vallecano | - | Athletic Club | 2:0 (2:0) | ||
| R | Athletic Club | - | Rayo Vallecano | 2:2 (1:0) | ||
| H: 28.10.2009 20:00 | R: 11.11.2009 21:00 | Sieger: Rayo Vallecano | ||||
| H | RC Celta | - | CD Tenerife | 2:1 (2:1) | ||
| R | CD Tenerife | - | RC Celta | 0:1 (0:0) | ||
| H: 28.10.2009 20:00 | R: 10.11.2009 20:00 | Sieger: RC Celta | ||||
| H | Real Murcia | - | Deportivo La Coruña | 0:1 (0:0) | ||
| R | Deportivo La Coruña | - | Real Murcia | 0:0 (0:0) | ||
| H: 28.10.2009 20:00 | R: 10.11.2009 20:00 | Sieger: Deportivo La Coruña | ||||
| H | Hércules CF | - | UD Almería | 2:1 (0:1) | ||
| R | UD Almería | - | Hércules CF | 0:1 (0:0) | ||
| H: 28.10.2009 20:00 | R: 12.11.2009 21:00 | Sieger: Hércules CF | ||||
| H | Getafe CF | - | Espanyol Barcelona | 2:0 (0:0) | ||
| R | Espanyol Barcelona | - | Getafe CF | 1:1 (1:1) | ||
| H: 28.10.2009 20:00 | R: 11.11.2009 20:00 | Sieger: Getafe CF | ||||
| H | CD Alcoyano | - | Valencia CF | 0:1 (0:1) | ||
| R | Valencia CF | - | CD Alcoyano | 2:2 (2:1) | ||
| H: 28.10.2009 20:00 | R: 10.11.2009 19:00 | Sieger: Valencia CF | ||||
| H | Cultural Leonesa | - | FC Barcelona | 0:2 (0:1) | ||
| R | FC Barcelona | - | Cultural Leonesa | 5:0 (0:0) | ||
| H: 28.10.2009 22:00 | R: 10.11.2009 22:00 | Sieger: FC Barcelona | ||||
| H | Real Valladolid | - | RCD Mallorca | 2:1 (1:1) | ||
| R | RCD Mallorca | - | Real Valladolid | 1:0 (1:0) | ||
| H: 29.10.2009 21:00 | R: 11.11.2009 21:00 | Sieger: RCD Mallorca | ||||
| H | UD Salamanca | - | Racing Santander | 1:0 (0:0) | ||
| R | Racing Santander | - | UD Salamanca | 4:1 (2:1) | ||
| H: 29.10.2009 21:00 | R: 11.11.2009 21:00 | Sieger: Racing Santander | ||||
| H | Unión Deportiva Puertollano | - | Villarreal CF | 1:1 (1:0) | ||
| R | Villarreal CF | - | Unión Deportiva Puertollano | 1:0 (0:0) | ||
| H: 29.10.2009 21:00 | R: 10.11.2009 19:00 | Sieger: Villarreal CF | ||||
