The Hornets will visit Sutton United if they beat the Tykes at the Camping World Community Stadium on Tuesday, November 14.
The U’s, who sit bottom of League Two, advanced to the second round courtesy of a 2-1 home win over National League outfit AFC Fylde on Saturday.
The draw was conducted live on ITV1 by former Wimbledon goalkeeper Dave Beasant and ex-England and Arsenal defender Anita Asante.
READ THIS: Dominic Di Paola hails ‘absolutely phenomenal’ Horsham performance as Barnsley are held in FA Cup barnstormer
Horsham manager Dominic Di Paola lauds FA Cup draw with Barnsley as ‘best performance in club’s history’
38 brilliant match and fan photos as never-say-die Horsham hold League One Barnsley in roller coaster first round tie
The 20 ties in the second round proper will take place across the weekend of Saturday, December 2, and the winning sides will take home £67,000 from the competition's prize fund.
Horsham were in the hat for the second round following their heroic 3-3 draw at Barnsley on Friday evening.
The Hornets have beaten Leatherhead, Marlow, Hanworth Villa and Dorking Wanderers in the previous qualifying rounds of this season’s competition.