For just the fourth time in the club's history, the Glovers take on Manchester United on Friday, January 26th (7.55pm) live on BBC One.
Town are the lowest ranked team left in the competition and will be looking to add another chapter to the club's illustrious giant-killing story.
Having reached round four for just the fourth time ever, Darren Way's side will now bid to defy the odds and secure a victory which would equal Yeovil's best ever run in the competition.
The Green and Whites will welcome a different looking United side to that of the last meeting in 2015 having reached the fourth step of the FA Cup (without a bye) for the first time since the 2004/05 season.
High profile names such as Paul Pogba, Marcus Rashford, Romelu Lukaku and Alexis Sanchez are all now part of the roster, as the Premier League giants head to Somerset.
Managed by Jose Mourinho, the Red Devils head to Huish Park second in the top flight behind rivals Manchester City.
Arriving earlier in the week from Arsenal, Chilean international Sanchez is in line to make his debut on Friday night in front of the sell out 9,000 crowd.
Looking to cause one of the biggest cup upsets of all time, Town will be without the services of Oscar Gobern (hamstring), James Bailey (knee) and Dan Alfei (knee), but are likely to welcome back Rhys Browne for the first time since November after a lengthy ankle injury.
Joint top scorer Otis Khan will serve the first of a five-match suspension whilst striker Marcus Barnes is still ruled out for more than a month.
Manchester United have an unblemished record against the Glovers.
In the three previous meetings, United have netted 13 goals without conceding, most recently recording a 2-0 win at Huish Park three years ago courtesy of Ander Herrera and Angel Di Maria.
In Town's only ever outing in the FA Cup fifth round in 1949, the Red Devils ran out 8-0 victors.
Paul Tierney (Lancashire) will be the man in the middle at Huish Park.
Taking charge of his 19th game of the season, it will be his first involving Yeovil since April 2016 having overseen the 2-1 defeat to Bristol Rovers at the Memorial Ground in which he awarded three yellow cards.
On Friday, Steve Child (Kent) and Sian Massey-Ellis (West Midlands) will be assistant referees with Keith Stroud (Hampshire) acting as fourth official.
The pre-match thoughts of manager Darren Way and players Sam Surridge, Jake Gray and Ryan Dickson can be viewed on iFollow Glovers.
If you’re unable to attend the game, live audio commentary is available via the iFollow Glovers service, with the monthly and season subscriptions offering full-match replays from Sunday.
A full match report will follow on YTFC.net at full time with post-match reaction from the manager and a member of the playing staff following soon after.
Alternatively, the match will be shown live on BBC One.