| Tuesday 6th October 2009 | |||
| Northern Second Round | |||
| Bradford City | 2-2 | Notts County | |
| (Bradford City won on penalties) | |||
| Bury | 2-1 | Tranmere Rovers | |
| Carlisle United | 4-2 | Macclesfield Town | |
| Chesterfield | 3-3 | Huddersfield Town | |
| (Chesterfield won on penalties) | |||
| Hartlepool United | 0-2 | Grimsby Town | |
| Leeds United | 2-1 | Darlington | |
| Port Vale | 3-1 | Stockport County | |
| Southern Second Round | |||
| Charlton Athletic | 4-1 | Barnet | |
| Exeter City | 1-1 | Swindon Town | |
| (Swindon Town won on penalties) | |||
| Gillingham | 0-1 | Norwich City | |
| Hereford United | 2-2 | Aldershot Town | |
| (Hereford United won on penalties) | |||
| Leyton Orient | 1-0 | Brighton and Hove A | |
| Milton Keynes Dons | 2-0 | Southend United | |
| Northampton Town | 2-1 | Bournemouth | |
| Southampton | 2-2 | Torquay United | |
| (Southampton won on penalties) | |||
» English Johnstone's Paint Trophy 2009-2010 : Results
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