Key: Win : Draw : Loss
| Date | Home | Away | Ref | ||
|---|---|---|---|---|---|
| 08/08/2009 | Yeovil | 2 | 0 | Tranmere | A Hall |
| 15/08/2009 | Tranmere | 4 | 2 | Gillingham | C Pawson |
| 18/08/2009 | Tranmere | 0 | 1 | Milton Keynes Dons | J Moss |
| 22/08/2009 | Leeds | 3 | 0 | Tranmere | E Ilderton |
| 29/08/2009 | Tranmere | 0 | 4 | Charlton | P Tierney |
| 05/09/2009 | Carlisle | 3 | 0 | Tranmere | C Boyeson |
| 12/09/2009 | Tranmere | 2 | 3 | Walsall | M Haywood |
| 19/09/2009 | Exeter | 2 | 1 | Tranmere | D Phillips |
| 26/09/2009 | Tranmere | 1 | 1 | Colchester | S Bratt |
| 29/09/2009 | Wycombe | 0 | 1 | Tranmere | F Graham |
| 03/10/2009 | Millwall | 5 | 0 | Tranmere | G Scott |
| 12/10/2009 | Tranmere | 0 | 1 | Stockport | R Booth |
| 17/10/2009 | Tranmere | 2 | 1 | Brighton | J Waugh |
| 24/10/2009 | Hartlepool | 1 | 0 | Tranmere | G Sutton |
| 31/10/2009 | Tranmere | 1 | 4 | Swindon | A Bates |
| 14/11/2009 | Norwich | 2 | 0 | Tranmere | D Deadman |
| 21/11/2009 | Leyton Orient | 2 | 1 | Tranmere | D Sheldrake |
| 24/11/2009 | Tranmere | 2 | 0 | Southend | D Webb |
| 01/12/2009 | Huddersfield | 3 | 3 | Tranmere | N Swarbrick |
| 05/12/2009 | Tranmere | 1 | 0 | Brentford | C Sarginson |
| 12/12/2009 | Southampton | 3 | 0 | Tranmere | D McDermid |
| 19/12/2009 | Tranmere | 2 | 0 | Bristol Rvs | J Singh |
| 28/12/2009 | Tranmere | 0 | 0 | Carlisle | P Gibbs |
| 20/01/2010 | Oldham | 0 | 0 | Tranmere | C Pawson |
| 23/01/2010 | Milton Keynes Dons | 1 | 0 | Tranmere | M Haywood |
| 26/01/2010 | Tranmere | 2 | 1 | Yeovil | G Salisbury |
| 30/01/2010 | Charlton | 1 | 1 | Tranmere | G Sutton |
| 06/02/2010 | Tranmere | 0 | 1 | Oldham | C Sarginson |
| 09/02/2010 | Gillingham | 0 | 1 | Tranmere | R East |
| 13/02/2010 | Southend | 1 | 1 | Tranmere | S Cook |
| 20/02/2010 | Tranmere | 2 | 1 | Leyton Orient | D Foster |
| 23/02/2010 | Tranmere | 0 | 2 | Huddersfield | A Penn |
| 06/03/2010 | Tranmere | 2 | 1 | Southampton | P Quinn |
| 09/03/2010 | Tranmere | 1 | 4 | Leeds | R Shoebridge |
| 13/03/2010 | Bristol Rvs | 0 | 0 | Tranmere | G Scott |
| 19/03/2010 | Tranmere | 0 | 0 | Hartlepool | K Evans |
| 27/03/2010 | Brighton | 3 | 0 | Tranmere | P Taylor |
| 02/04/2010 | Tranmere | 3 | 1 | Norwich | E Ilderton |
| 05/04/2010 | Swindon | 3 | 0 | Tranmere | O Langford |
| 10/04/2010 | Walsall | 2 | 1 | Tranmere | J Waugh |
| 13/04/2010 | Tranmere | 0 | 3 | Wycombe | A Haines |
| 17/04/2010 | Tranmere | 3 | 1 | Exeter | G Eltringham |
| 20/04/2010 | Brentford | 2 | 1 | Tranmere | D Phillips |
| 24/04/2010 | Colchester | 1 | 1 | Tranmere | P Crossley |
| 01/05/2010 | Tranmere | 2 | 0 | Millwall | N Swarbrick |
| 08/05/2010 | Stockport | 0 | 3 | Tranmere | T Kettle |