Two of the biggest names in the tournament go head-to-head in Franceville, after unconvincing wins in their opening games.
Ghana are without captain John Mensah, after the defender was sent off in their tight 1-0 win over Botswana.
The Black Stars are also waiting on the fitness of Asamoah Gyan, after he limped out of training on Thursday.
Mali's Seydou Keita has called for team unity as the Eagles seek a first quarter-final berth in eight years.
Ghana coach Goran Stevanovic will be forced to field a relatively inexperienced centre-back pairing in his team's second Group D game.
Mensah's namesake Jonathan Mensah is likely to line up in his place, alongside John Boye - who made his Nations Cup debut against Botswana - tipped to keep his place, as another regular centre-half, Isaac Vorsah, completes a two-match suspension.
"It's difficult to replace a player like Mensah," Stevanovic said.
"But I have told the team that I would need all of them for this tournament and so I have 20 players who could replace him against Mali."
Stvanovic will await further news on Gyan, who limped out of his training with a suspected ankle, but he is expected to recover in time for Saturday's game.
Meanwhile, Keita - who captained the Eagles to the last four at the 2002 Nations Cup - said Mali are driven to beat the Ghanaians.
"Ghana remain the favourites in our group, but the Black Stars are not unbeatable," Keita said, who was emotional after a hard-fought 1-0 win over neighbours Guinea on Tuesday.
"The objective is to play to our potentials and get a positive result on Saturday.
"We have not achieved anything yet and in football.
"Things can happen so fast and so our second game against Ghana will be a lot more decisive."
Keita has also admitted to feeling the pressure himself, as he leads a youthful Mali team at the finals.
"Personally, I have been under a lot of pressure but I will continue to assume my responsibilities," said Keita, who spent several minutes giving thanks to Allah immediately after Tuesday's win.