Juventus has the edge on away goals thanks to Ronaldo's 125th Champions League goal in a 1-1 draw in Amsterdam, but Ajax's David Neres ensured the teams will start on a knife-edge in Turin in a repeat of the 1973 and 1996 finals, AFP reported.
Juventus has put its bid to wrap up an eighth consecutive Serie A title on the back burner as it focuses on its European ambitions.
Massimiliano Allegri lined out with a young squad against lowly SPAL on Saturday and the champion fell to a 2-1 defeat when a point would have sufficed for another Scudetto.
"The Scudetto will come sooner or later," said Allegri, whose side hold a 17-point lead on second-placed Napoli with six games left.
"If we had put all our starters in, it would have been easier to win it, but there is the goal to obtain on Tuesday."
Juventus has won the Champions League twice, most recently in 1996 when it beat Ajax in the final.
Four-time winner Ajax won its third consecutive title in 1973 at the expense of Juventus, last lifting the trophy in 1995.
Allegri said he was prepared for "a difficult match against a very impressive team".
Ronaldo was rested at the weekend along with regulars Leonardo Bonucci, Blaise Matuidi, Alex Sandro, Mario Mandzukic and Miralem Pjanic.
Teenager Moise Kean's fine form continued with the 19-year-old scoring his sixth goal in as many games.
Captain Giorgio Chiellini, however, remains a doubt with the 34-year-old defender recovering from a calf injury, with Daniele Rugani a possible replacement.
But Ronaldo remains Juve's key weapon.
The 34-year-old was signed for €100 million ($117 million) last summer, and pocketed an annual €31 million salary to help Juventus win a competition it has finished runner-up in a record seven times, most recently in 2015 and 2017.
Ronaldo has played a leading role in all of his five previous European triumphs, including with the tie-clinching, 97th-minute penalty for Real Madrid against Juventus last season in the quarters.
He has scored five goals this campaign, one against Manchester United and a hat-trick which rescued Juventus in the last 16, second leg against Atletico, before returning from injury to score against Ajax.