The hosts saw off Barnet to seal their place in the round two, while the visitors got their FA Cup journey up and running with a win over Horsham.
Match preview
Exeter are competing in the second round after they staged a dramatic fightback from 3-2 down to get the better of National League Barnet earlier this month.
Josh Magennis equalised in the 85th minute, before Jay Bird and Kami Doyle scored in stoppage time to clinch a 5-3 victory, sending Exeter into the second round for the third time in four seasons.
The Grecians followed that win with a narrow 1-0 victory over Charlton Athletic, but they have since experienced an underwhelming period of form, having endured a three-game winless run without troubling the scorers.
After playing out a goalless draw with Lincoln City, the Grecians went on to lose 3-0 to Wrexham and 2-0 to Birmingham City, leaving them in 11th position in League One and four points adrift of the playoffs.
Exeter will put their league campaign on hold as they look to reach the FA Cup third round for the first time since 2020-21 when they prevailed 3-2 in a round two meeting with Gillingham.
Chesterfield are sitting just one point outside of the League Two playoffs after winning six, drawing seven and losing four of their 17 league matches this season.
Paul Cook's side recently bounced back from consecutive league defeats to claim a narrow 1-0 victory over Barrow, Jamie Grimes scoring an 89th-minute winner at the SMH Group Stadium.
They will now head on their travels for Saturday's FA Cup clash, with the Spireites set to compete in the second round for the fifth straight season after beating seventh-tier Horsham 3-1 in the previous round.
Chesterfield have enjoyed success in their recent appearances at this stage of the FA Cup, having prevailed in each of their previous three second-round ties.
The visitors may be able to take some confidence from the fact they have won three of their previous five away matches, scoring at least three goals in each of those victories.
Exeter City FA Cup form:
W
Exeter City form (all competitions):
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D
L
L
Chesterfield FA Cup form:
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Chesterfield form (all competitions):
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L
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W
Team News
Exeter's Johnly Yfeko and Patrick Jones are both nursing hamstring injuries, while Yanic Wildschut remains sidelined with a long-term knee injury.
Midfielder Reece Cole has been absent for the last four matches, making him an obvious doubt for Saturday's cup tie.
The hosts will also have to assess the fitness of Millenic Alli, who sat out the recent defeat to Birmingham due to an injury issue.
As for the visitors, they are expected to be without the services of Ryheem Sheckleford, Ash Palmer, John Fleck and Chey Dunkley.
Grimes could be brought into the starting lineup after featuring as a half-time substitute in the recent win over Barrow.
Cook may also take the opportunity to offer starting spots to the likes of Michael Jacobs, Armando Dobra, Ryan Colclough and Will Grigg.
Exeter City possible starting lineup:
Whitworth; Sweeney, Crama, Francis; Niskanen, McMillan, Woods, Harper; Aitchison; Bird, Magennis
Chesterfield possible starting lineup:
Boot; Tanton, Williams, Grimes, Gordon; Oldaker, Naylor; Jacobs, Dobra, Colclough; Grigg