The Tykes are hoping to avoid consecutive first-round exits, while the Valiants are looking to progress to round two for the second straight season.
Match preview
Port Vale will enter the FA Cup tie as the League Two leaders after racking up 30 points from 14 games to establish a four-point lead at the top of the table.
Impressively, the Valiants have won each of their last five matches, including a narrow win in last Saturday's entertaining clash with Wimbledon.
Brandon Cover, Rico Richards and Jayden Stockley all got on the scoresheet to fire Port Vale to a 3-2 victory, extending their unbeaten run to 11 matches in all competitions.
The Valiants have also won four of their last five home matches, and they will hope that run of results can help them end a four-game winless home run in the FA Cup.
They will also be looking to beat Barnsley for the first time since November 2022 after losing each of their last three head-to-head meetings.
Barnsley are sitting in eighth place in the League One table after winning six, drawing four and losing three of their 13 matches.
The Tykes have put together a three-game unbeaten run in the league, having claimed wins against Blackpool and Shrewsbury Town on either side of a draw with Charlton Athletic.
In contrast to their recent league form, Barnsley experienced disappointment in the midweek EFL Trophy clash against Doncaster Rovers, Aiden Marsh's effort proving to be a mere consolation in a 3-1 defeat.
After losing at Oakwell on Tuesday, the Tykes will now head on their travels with hopes of recording a third consecutive away win.
They will hope to produce another strong away performance as they look to avoid a second straight first-round exit after being knocked out by non-league Horsham following a replay in last season's competition.
Port Vale form (all competitions):
D
W
W
W
W
W
Barnsley form (all competitions):
L
L
W
D
W
L
Team News
Port Vale boss Darren Moore will be unable to call upon the injured quartet of Mitch Clark, Sam Hart, George Byers and Kyle John.
Moore could be tempted to stick with the same lineup that started the narrow 3-2 win over Wimbledon last weekend.
Stockley is poised to lead the line for the Valiants, with the striker looking to add to his four-goal tally for the season.
As for the visitors, Matty Wolfe remains unavailable due to injury, while Sam Cosgrove is unlikely to be ready despite making progress his his recovery.
Former Port Vale manager Darrell Clarke has revealed that Adam Phillips is closing in on a return to action following an ankle injury, but a final decision is yet to be made on his involvement in Saturday's cup tie.
Max Watters is set to spearhead the Barnsley attack after finding the net in his first two appearances since returning from injury.
Port Vale possible starting lineup:
Ripley; Sang, Debrah, Hall; Cover, Croasdale, Garrity, Shorrock; Richards, Chislett; Stockley
Barnsley possible starting lineup:
Killip; De Gevigney, Pines, Roberts; O'Keeffe, Connell, Russell, Gent; Keillor-Dunn, Humphrys; Watters
We say: Port Vale 2-2 Barnsley
Barnsley may be competing in the league above Port Vale, but we think they could experience a tough outing at Vale Park on Saturday.