As this was the last full FA Cup competition before the Second World War, Portsmouth held the trophy until the end of the 1945–46 season.
If scores were level after 90 minutes had been played, a replay would take place at the stadium of the second-named team later the same week.
The 25 winners from the fourth qualifying round were North Shields, Horden Colliery Welfare, Lancaster City, Stalybridge Celtic, Chorley, Workington, Runcorn, Scunthorpe & Lindsey United, Gainsborough Trinity, Wellington Town, Kidderminster Harriers, Peterborough United, Colchester United, Lowestoft Town, Chelmsford City, Apsley (later to be known as Hemel Hempstead Town), Hayes, Corinthian, Folkestone, Guildford City, Walthamstow Avenue, Tunbridge Wells Rangers, Street, Cheltenham Town and Yeovil & Petter's United.
Hayes was the only club to progress from the extra preliminary round to the first round proper, defeating Wycombe Wanderers, Yiewsley, Chesham United, Banbury Spencer, Southall and Newport (IOW) to secure their place in the main draw.
However, Chelmsford City was the most successful non-league club this season, moving from the preliminary round to the fourth round proper and defeating Barking, Ford Sports (Dagenham), Crittall Athletic, Romford, Dulwich Hamlet, Kidderminster Harriers and Football League sides Darlington and Southampton in their history-making run.
The 44 First and Second Division clubs entered the competition at this stage along with Barnsley, York City and Notts County.
Chelmsford City was the last non-league club remaining in the competition, participating in their ninth round of this tournament.