| Matches by round |
| 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 |
| Match list: | By date Latest | Player stats: |
| R | Date | Home | Score | Away | |
| 1 | 16/08/2009 | Tranmere Rovers | 5 - 0 | Scunthorpe Unit | |
| 2 | 23/08/2009 | Middlesbrough | 2 - 3 | Tranmere Rovers | |
| 3 | 30/08/2009 | South Durham | 3 - 0 | Tranmere Rovers | |
| 4 | 06/09/2009 | Tranmere Rovers | 2 - 0 | Liverpool Feds | |
| 5 | 13/09/2009 | Rochdale | 5 - 1 | Tranmere Rovers | |
| 7 | 27/09/2009 | Bradford City | 7 - 3 | Tranmere Rovers | |
| 8 | 04/10/2009 | Tranmere Rovers | 2 - 0 | Wakefield | |
| 10 | 18/10/2009 | Tranmere Rovers | 3 - 3 | Stockport Co | |
| 11 | 25/10/2009 | Tranmere Rovers | 9 - 0 | Darlington | |
| 15 | 13/12/2009 | Tranmere Rovers | 0 - 2 | Rochdale | |
| 19 | 14/02/2010 | Scunthorpe Unit | 2 - 0 | Tranmere Rovers | |
| 23 | 14/03/2010 | Tranmere Rovers | - | Bradford City | |
| 24 | 21/03/2010 | Salford | - | Tranmere Rovers | |
| 25 | 28/03/2010 | Blackpool Wren | - | Tranmere Rovers | |
| 26 | 04/04/2010 | Darlington | - | Tranmere Rovers | |
| 27 | 11/04/2010 | Tranmere Rovers | - | Middlesbrough | |
| 28 | 18/04/2010 | Tranmere Rovers | - | Blackpool Wren | |
| 29 | 25/04/2010 | Stockport Co | - | Tranmere Rovers | |
| 30 | 02/05/2010 | Tranmere Rovers | - | Salford | |
| 31 | 09/05/2010 | Liverpool Feds | - | Tranmere Rovers |