The Valiants are making their first appearance in the third round since the 2009-10 season, while the visitors are enjoying their best-ever run in the competition.
Port Vale have enjoyed an encouraging start to their second straight League One season, collecting 17 points from their opening nine matches to occupy fourth position in the table.
The Valiants head into the midweek encounter off the back of a 1-1 draw with Cambridge United, where Connor Ripley saved Fejiri Okenabirhie's 90th-minute penalty to ensure that his side returned home with a point.
As a result of Saturday's away draw, Port Vale have now only lost one of their last 11 competitive matches, including two positive results in the EFL Cup.
After beating Fleetwood Town 3-2 in round one, the Valiants held their nerve from the penalty spot to win a shootout against Crewe Alexandra to book their place in the third round of the competition for the first time in 14 years.
With Port Vale targeting their first last-16 appearance since the 2006-07 season, they may draw inspiration from the fact that they won 2-0 in their one and only competitive home meeting with Sutton in March 2022.
Sutton, meanwhile, have endured a tough start to the League Two season, having collected just four points from their nine matches to leave them bottom of the table.
They began the campaign with an impressive 5-1 victory over Notts County, but instead of building upon that result, they suffered seven consecutive league defeats before they ended their losing run with a 1-1 draw against MK Dons at the weekend.
Defender Omar Sowunmi scored his first goal for the club to give Sutton a half-time lead on Saturday, but Alex Gilbey restored parity within eight minutes of the restart to ensure that the spoils were shared.
While Matt Gray's side have struggled in the league, the EFL Cup has offered some much-needed respite after securing their place in the third round with a penalty-shootout win over Cambridge United and a 1-0 away victory over Wycombe Wanderers.
Prior to this season, Sutton had failed to make it past the first round in their previous two appearances in the EFL Cup, but they now have the chance to secure their place in the round of 16, as they bid to eliminate League One opposition for the third time in this season's competition.
After featuring as a substitute in the last three matches, Lewis Cass could be brought into the backline for Tuesday's contest.
Thomas Sang is also in contention to start against Sutton after coming off the bench at half time of Saturday's draw with Cambridge.
As for the visitors, Scott Kashket is set for an extended spell on the sidelines after sustaining an anterior cruciate ligament injury in last week's EFL Trophy defeat to Aston Villa Under-21s.
Sutton showed signs of improvement in their 1-1 draw, and Gray could be tempted to stick with the same starting lineup for Tuesday's cup tie.
After making his first league appearance for the club on Saturday, Reading loanee Dean Bouzanis will be looking to keep Sutton's second competitive clean sheet of the season.
Port Vale possible starting lineup:
Ripley; Cass, Smith, Iacovitti; Sang, Arblaster, Ojo, Plant; Garrity, Chislett; Wilson
Sutton United possible starting lineup:
Bouzanis; Kizzi, Sowumi, John, Hart; Coley, Clay, N'Guessan, Patrick; Smith, O'Brien
We say: Port Vale 2-1 Sutton United
Although Sutton have knocked out two teams in League One to book their place in third round, we think that they will be unable to repeat that feat on Tuesday.
We believe that the Valiants will prove too strong for a Sutton side who have failed to win any of their last five competitive games.
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