Southend United visit the Kassam Stadium in the Johnstone’s Paint Trophy Southern semi-finals.
The winners will go through to the two-legged area final next month – the final hurdle before the showpiece at Wembley in April.
Wins over Swindon, Cheltenham and Plymouth have ensured this is already United’s best season in the competition.
But with a rare chance to play at the national stadium on the horizon, Wilder is determined to make the most of it.
“It’s going to be a tight tie, but one we can certainly win,” United’s manager said. “It’s a massive opportunity for the players and the supporters.
“We’ve worked hard to get through some difficult ties and we’ll have to do it again, but it really is game on for us and we have to grab this chance.”
The prospect of three games in eight days may be too much for the fit-again Michael Duberry and, with one eye on Saturday’s league trip to Bradford, Michael Raynes could be preferred to the veteran at centre half.
Sean Rigg will definitely start after coming on as a substitute in Saturday’s 3-0 FA Cup defeat by Sheffield United.
The winger may be asked to play in a role just behind top scorer Tom Craddock, with James Constable suspended, and Justin Richards, who started against the Blades, cup-tied.
Fellow new signing Lewis Montrose is also unavailable, but fit-again Liam Davis is back in contention.
Southend lost their league game here 2-0 in August, but had won their previous six meetings with the U’s, including a third-round tie in this competition last season.
United midfielder Simon Heslop feels a desire to make up for past setbacks, along with thoughts of reaching the final, will act as powerful incentives.
He said: “We shouldn’t have any problems motivating ourselves. We know all about Southend – they’ve had a few wins against us in the past, so that will drive us on.”
Southend striker Britt Assombalonga will have a late fitness test after missing Saturday’s 2-2 draw with Brentford in the FA Cup.
Oxford Utd: from Clarke, Batt, Raynes, Wright, O’Brien, Potter, Heslop, Leven, Capaldi, Rigg, Craddock, Davis, Duberry, Worley, Parker, Marsh.
Southend Utd: from Smith, Clohessy, Cresswell, Phillips, Prosser, Woodyard, Hurst, Martin, Timlin, Tomlin, Assombalonga, Corr, Benyon, Eastwood, Straker, Bentley, Spicer, Barker.
OXFORD United’s League Two fixture against Fleetwood, postponed before Christmas because of waterlogging at the Kassam Stadium, has been re-arranged for Tuesday, February 12.