| Tuesday, September 29, 2009 | |||||
| Time | Home | Away | VENUE | ||
| 19:45 UK | Barnet | v | Dagenham & Redbridge | Underhill Stadium | |
| 19:45 UK | Burton Albion | v | Macclesfield Town | Pirelli Stadium | |
| 19:45 UK | Cheltenham Town | v | Shrewsbury Town | Whaddon Road Ground | |
| 19:45 UK | Crewe Alexandra | v | Bury | The Alexandra Stadium | |
| 19:45 UK | Darlington | v | Rochdale | The Balfour Webnet Darlington Arena | |
| 19:45 UK | Hereford United | v | AFC Bournemouth | Edgar Street | |
| 19:45 UK | Lincoln City | v | Notts County | Sincil Bank | |
| 19:45 UK | Morecambe | v | Bradford City | Christie Park | |
| 19:45 UK | Northampton Town | v | Rotherham United | Sixfields Stadium | |
| 19:45 UK | Port Vale | v | Accrington Stanley | Vale Park | |
| 19:45 UK | Torquay United | v | Aldershot Town | Plainmoor Ground | |
