Chesterfield will make the short trip to Alfreton Town on Tuesday night (kick-off 7.30pm).
The Spireites will take a full senior squad to the Impact Arena for the match.
Manager Paul Cook is likely to use the game to take another look at one or two triallists who are keen to impress.
Alfreton, who are managed by former Spireites boss Nicky Law, are in the Conference Premier division.
Admission to the game costs £10 for adults, £5 for concessions and £1 for under-12s who are accompanied by a paying adult.
Meanwhile, Tuesday night will also see a Chesterfield XI featuring players with first team experience in action at Sheffield FC (kick-off 7.30pm).
*Please note that plans to take the Sky Bet League 2 trophy to Sheffield FC have now had to be cancelled.