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| Date | Opposition | H/A | Res | Score | Competition | Scorers | Att | Report |
| 07|07|2009 | Dereham Town | Away | W | 2 - 0 | Friendly | Barker, Bokare | | match report | |
| 11|07|2009 | West Bromwich Albion | Home | W | 1 - 0 | Friendly | Barker | 1058 | match report | |
| 14|07|2009 | Northampton Town | Home | D | 1 - 1 | Friendly | Simpson | 341 | match report | |
| 23|07|2009 | Bury Town | Away | D | 0 - 0 | Friendly | | 188 | match report | |
| 28|07|2009 | Colchester United | Home | L | 0 - 1 | Friendly | | | | |
| 03|08|2009 | Cambridge United | Away | W | 2 - 1 | CPC | Hammond, Simpson | 886 | match report | |
| 08|08|2009 | Gateshead | Away | W | 3 - 0 | League | Wright 2, Hudson-Odoi | 681 | match report | |
| 11|08|2009 | Grays Athletic | Home | D | 0 - 0 | League | | 816 | match report | |
| 15|08|2009 | Oxford United | Home | L | 3 - 4 | League | | 1433 | match report | |
| 18|08|2009 | Altrincham | Away | L | 1 - 2 | League | Simpson | 698 | match report | |
| 22|08|2009 | Salisbury City | Home | W | 2 - 0 | League | Frew, Wright | 684 | match report | |
| 29|08|2009 | York City | Away | L | 1 - 3 | League | Wright | 1944 | match report | |
| 31|08|2009 | Stevenage Borough | Home | L | 0 - 2 | League | | 1159 | match report | |
| 05|09|2009 | Chester City | Away | L | 0 - 2 | League | | 1171 | match report | |
| 08|09|2009 | Crawley Town | Away | L | 0 - 2 | League | | 798 | match report | |
| 12|09|2009 | Kidderminster Harriers | Home | D | 1 - 1 | League | Hammond | 774 | match report | |
| 19|09|2009 | Hayes & Yeading United | Away | W | 2 - 0 | League | Knight-Percival, Simpson | | match report | |
| 22|09|2009 | Ebbsfleet United | Home | W | 1 - 0 | League | Simpson | 647 | match report | |
| 26|09|2009 | AFC Wimbledon | Away | L | 0 - 4 | League | | 3392 | match report | |
| 29|09|2009 | Eastbourne Borough | Home | W | 2 - 0 | League | Knight-Percival 2 | 633 | match report | |
| 04|10|2009 | Cambridge United | Home | D | 1 - 1 | League | Langston | 2371 | match report | |
| 10|10|2009 | Tamworth | Away | W | 3 - 1 | League | Knight-Percival 2, Simpson | 1090 | match report | |
| 17|10|2009 | Forest Green Rovers | Home | W | 5 - 2 | League | Knight-Percival, Barker, Simpson, Wright 2 | 823 | match report | |
| 24|10|2009 | Hinckley United | Away | L | 1 - 2 | FA Cup 4Q | Oyebanjo | 778 | match report | |
| 31|10|2009 | Mansfield Town | Away | D | 1 - 1 | League | Simpson (p) | 3162 | match report | |
| 14|11|2009 | Rushden & Diamonds | Away | L | 1 - 2 | League | Tidswell | 1126 | match report | |
| 21|11|2009 | Wrexham | Home | D | 0 - 0 | League | | 946 | match report | |
| 24|11|2009 | Eastbourne Borough | Away | D | 1 - 1 | League | Hudson-Odoi | 703 | match report | |
| 01|12|2009 | Grays Athletic | Away | W | 1 - 0 | League | Smith | 367 | match report | |
| 05|12|2009 | Altrincham | Home | D | 0 - 0 | League | | 676 | match report | |
| 28|12|2009 | Kettering Town | Home | W | 1 - 0 | League | Tann | 716 | match report | |
| 19|01|2010 | Maidstone United | Away | W | 3 - 0 | FA Trophy R1 | Wright (pen), Tidswell, Tann (Histon expelled due to ineligible player) | 238 | match report | |
| 23|01|2010 | Salisbury City | Away | L | 0 - 3 | League | | 758 | match report | |
| 27|01|2010 | Luton Town | Home | L | 0 - 2 | League | | 1543 | match report | |
| 02|02|2010 | York City | Home | D | 1 - 1 | League | Hudson-Odoi | 487 | match report | |
| 06|02|2010 | Hayes & Yeading United | Home | D | 3 - 3 | League | Wright (pen), Langston, Smith | 617 | match report | |
| 09|02|2010 | Ebbsfleet United | Away | W | 1 - 0 | League | Wright | 731 | match report | |
| 13|02|2010 | Oxford United | Away | L | 0 - 2 | League | | 5365 | match report | |
| 27|02|2010 | Kidderminster Harriers | Away | L | 0 - 3 | League | | 1293 | match report | |
| 03|03|2010 | Rushden & Diamonds | Home | L | 0 - 1 | League | | 583 | match report | |
| 06|03|2010 | Gateshead | Home | D | 0 - 0 | League | | 645 | match report | |
| 09|03|2010 | Tamworth | Home | W | 1 - 0 | League | Smith | 350 | match report | |
| 13|03|2010 | Cambridge United | Away | L | 1 - 2 | League | Pugh | 4417 | match report | |
| 16|03|2010 | Stevenage Borough | Away | L | 0 - 1 | League | | 2407 | match report | |
| 20|03|2010 | Mansfield Town | Home | L | 0 - 5 | League | | 662 | match report | |
| 27|03|2010 | Wrexham | Away | L | 0 - 3 | League | | 2335 | match report | |
| 29|03|2010 | Barrow | Away | D | 0 - 0 | League | | 994 | match report | |
| 03|04|2010 | AFC Wimbledon | Home | L | 1 - 3 | League | Johnson o.g. | 867 | match report | |
| 05|04|2010 | Kettering Town | Away | D | 1 - 1 | League | Smith | 800 | match report | |
| 10|04|2010 | Crawley Town | Home | L | 0 - 1 | League | | 499 | match report | |
| 13|04|2010 | Luton Town | Away | L | 3 - 6 | League | Sparkes, Knight-Percival, Southam | 7083 | match report | |
| 17|04|2010 | Forest Green Rovers | Away | L | 0 - 2 | League | | 1204 | match report | |
| 24|04|2010 | Barrow | Home | D | 2 - 2 | League | Southam (pen), Jones o.g. | 753 | match report | |
On 18 February 2010 Chester City were expelled from the Football Conference, their record for 2009/10 was expunged. This means that the match at The Deva Stadium on 5 September 2009 is no longer in the record books, it is shown above for completeness.
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