The Spanish giants took a giant stride towards the Champions League final four by winning the first leg of their quarter-final tie 3-0 last week.
Madrid were hammered 4-1 by a Robert Lewandowski-inspired Dortmund in the semi-final of the competition almost 12 months ago.
Robert Lewandowski scored all four goals as the Blancos were swept aside at Signal Iduna Park and a 2-0 victory for the Liga side in the second leg at the Santiago Bernabeu was in vain as Jurgen Klopp’s side hung on to secure their place in the final.
Bundesliga rivals Bayern Munich then consigned the Schwarzgelben to a 2-1 defeat in the Wembley final though it looks unlikely they will go one better this season.
Dortmund’s chances of pulling off a stunning comeback are enhanced by the return of Bayern-bound striker Lewandowski for the second leg.
The prolific frontman missed the first leg due to suspension, but he is set to lead the line against Carlo Ancelotti’s treble-chasing side.
Lewandowski was on target as Dortmund warmed up for their crunch clash with Real by beating Wolfsburg 2-1 on Saturday, moving them closer to Champions League football next season.
Ivica Olic gave Wolfsburg a half-time lead at Dortmund on Saturday, but Klopp masterminded a second-half fightback by bringing on Milos Jojic and Erik Durm at the break, with Kevin Grosskreutz moving into midfield.
The Dortmund coach could stick with that winning formula against Madrid, who also come into the game on the back of a victory.
Ancelotti’s side hammered Real Sociedad 4-0 on Saturday to stay in the hunt for a treble of La Liga, Copa del Rey and Champions League, but their win came at a cost as Gareth Bale suffered a knee wound when he was caught by defender Markel Bergara.
The Wales international may be forced out of the second leg, but Ballon d’Or-winner Cristiano Ronaldo could be back in the side after returning to training. The Portugal captain missed was rested on Saturday due to a knee problem of his own, but looks set to feature in Dortmund as he attempts to add to his record-equalling tally of 13 Champions League goals for the season.
Ronaldo struck in the first leg to equal the record set by Lionel Messi two years ago and it would be no surprise to see him strike again.
Dortmund will be without midfielder Sebastian Kehl due to suspension, while goalkeeper Roman Weidenfeller is a doubt due to an arm injury alongside a hefty list of long-term absentees that includes Ilkay Gundogan, Marcel Schmelzer and Sven Bender.
Kick-off time is 18:45 GMT.