The home team recorded a 2-1 victory over Crawley Town to progress to this stage, while AFC Wimbledon were 2-0 winners over Oxford United in the last round of the tournament.
Match preview
Peterborough will bring a four-game unbeaten run into Tuesday's contest, including a goalless draw against Lincoln City in League One on Saturday afternoon.
Darren Ferguson's side are third in the League One table, boasting 56 points from their 28 matches, which has left them just three points behind the leaders Portsmouth with a game in hand.
The Posh are focusing on the league, but they are bidding to win the EFL Trophy for the second time in their history, having also secured the title in 2013-14.
Peterborough finished top of Group D, which included Colchester United, Tottenham Hotspur Under-21s and Cambridge United, before recording a 3-0 victory over Arsenal Under-21s in the last-32 stage of the Southern section.
Ferguson's team then beat Crawley 2-1 in the last-16 stage of the Southern section last time out, and their reward for winning this match would be a semi-final against either Wycombe Wanderers or Brighton & Hove Albion Under-21s.
AFC Wimbledon, meanwhile, will enter this match off the back of a 2-1 victory over Mansfield Town on Saturday, with the result leaving them in eighth position in the League Two table.
Johnnie Jackson's side are only two points outside of the playoff positions in the fourth tier of English football, which is an impressive achievement considering that they were 21st last season.
The Dons recorded a 2-0 victory over Oxford United in the last-16 stage of the Southern section, which followed a 5-2 success over Portsmouth in the round of 32.
AFC Wimbledon only managed one win in their three games in the group stage of the competition, but they finished second behind Wycombe in order to progress to the knockout stage and are now just one victory away from being present in the final four of the Southern section.
Peterborough recorded a 3-0 victory when these two sides last locked horns in February 2021, but AFC Wimbledon have won two of their last three encounters.
Peterborough United EFL Trophy form:
W
W
L
W
W
Peterborough United form (all competitions):
W
L
W
W
W
D
AFC Wimbledon EFL Trophy form:
W
W
L
W
W
AFC Wimbledon form (all competitions):
D
L
W
L
L
W
Team News
Peterborough head coach Ferguson could name the same side that took to the field for the first whistle against Crawley in the last round of the competition on January 23.
As a result, there are set to be changes from the XI that started the league encounter with Lincoln on Saturday, with Emmanuel Fernandez, Romoney Crichlow-Noble and Malik Mothersille among those expected to start.
Ephron Mason-Clark is the team's leading goalscorer this season with 14, and the 24-year-old is again set to feature in the final third of the field on Tuesday night.
As for AFC Wimbledon, head coach Jackson will make changes from the side that took to the field for the first whistle against Mansfield in the league on Saturday.
Nikola Tzanev is set to feature between the sticks, while Harry Pell is also set to be introduced in an advanced area for the League Two outfit.
James Tilley has scored 12 times in all competitions this season, including a brace against Oxford United in the previous round, and he will again feature in an advanced area on Tuesday night.
Peterborough United possible starting lineup:
Steer; Fernandez, Knight, Crichlow-Noble, Burrows; Fuchs, Collins; Ajiboye, De Havilland, Mason-Clark; Mothersille
AFC Wimbledon possible starting lineup:
Tzanev; Ogundere, Kalambayi, Johnson, Brown; Reeves, Ball; Tilley, Pell, Sasu; Davison
We say: Peterborough United 2-0 AFC Wimbledon
AFC Wimbledon are capable of picking up a positive result on Tuesday, but it is a huge ask against a talented Peterborough outfit. Ferguson's side are the favourites to progress to the semi-finals, and we are expecting them to triumph against their League Two opponents.