The away side beat Wolverhampton Wanderers Under-21s on penalties in the last round to advance, while Bolton recorded a 3-2 victory over Barrow to progress to this stage of the competition.
Match preview
Bolton suffered a demoralising defeat to Shrewsbury Town in League One on Saturday, conceding twice in the final 11 minutes of the contest in Shropshire to suffer a 3-2 loss.
Ian Evatt's side led 2-1 until the 79th minute of the match, when Christian Saydee levelled, and Chey Dunkley then netted the winner for the home side in the 92nd minute.
The loss has left Bolton in fifth spot in the table, boasting 32 points from their 20 matches, with the Trotters looking to challenge for a return to the Championship this term.
Bolton finished top of Group B in the Northern Section, picking up seven points in the process, to advance to the next round, where they recorded a 3-2 victory over Barrow to progress to the quarter-finals.
The Trotters were knocked out of the FA Cup by Barnsley at the start of November, so this competition has taken on added importance, although the league remains the priority competition for the club.
Man United Under-21s, meanwhile, overcame Wolves Under-21s on penalties in the last round of the competition following a goalless draw to advance to this stage.
Marc Jurado, Charlie McNeill, Manni Norkett and Kobbie Mainoo all scored during the shootout to send Mark Dempsey's side into the knockout round of the competition.
A number of the players have been away in Spain with the first team for their mid-season training camp, which included friendlies against Cadiz and Real Betis.
However, there is no reason, apart from injury, why the players should not be able to feature in this contest, so the Red Devils are likely to arrive with a strong squad on Tuesday night.
Man United Under-21s finished top of Group G of the Northern Section ahead of Barrow, with the team picking up seven points to progress to the round of 32.
Bolton Wanderers EFL Trophy form:
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D
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Bolton Wanderers form (all competitions):
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Manchester United Under-21s EFL Trophy form:
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Manchester United Under-21s form (all competitions):
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L
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Team News
Bolton boss Evatt will make changes to the side that started against Shrewsbury in the league on Saturday, with Joel Dixon likely to earn a start between the sticks.
Jon Dadi Bodvarsson should also feature in the final third of the field, with the striker on the scoresheet against Barrow in the last round.
However, Kyle Dempsey is set to miss out for the Trotters on Tuesday night through injury.
As for Man United Under-21s, head coach Dempsey will need to assess the fitness of the players who have been away with the first team in Spain.
Teden Mengi was taken off on a stretcher against Real Betis, though, with the centre-back now facing a period on the sidelines through injury.
Jurado, Mainoo and Charlie Savage should among those to feature for the Red Devils, while Tom Huddlestone, who is a player-coach for the Under-21s, could start in central midfield.
Bolton Wanderers possible starting lineup:
Dixon; Toal, Ricardo, Iredale; Sadlier, Williams, Sheehan, John; Charles, Kachunga, Bodvarsson
Manchester United Under-21s possible starting lineup:
Bishop; Jurado, Bennett, Hardley, Murray; Mainoo, Huddlestone, Savage; Emeran, Shoretire, Gore
We say: Bolton Wanderers 2-1 Manchester United Under-21s
We are expecting this to be a very competitive match, and both sides will fancy their chances of causing issues in the final third of the field. Bolton will have more experience, though, and we just have a feeling that the League One side will navigate their way into the next round of the competition courtesy of a narrow win.