Tranmere Rovers, quarter-finalists in 2004 but now a non-league side for the first time in 94 years, host fellow National League side Lincoln City.
The lowest-ranked sides left, ninth-tier pair Sporting Khalsa and Hoddesdon Town, will play FC United or Buxton, and Didcot Town respectively - provided they come through their replays.
Winners at this stage receive £12,500.
Midland League Premier Division Sporting Khalsa drew with Spalding United on Saturday. Hoddesdon, of the Spartan South Midlands League Premier Division, drew with Brentwood Town.
Northern Premier League Salford City, owed by Class of '92 quintet Ryan Giggs, Paul Scholes, Nicky Butt and the Neville brothers, Gary and Phil, who have a combined total of 16 FA Cup winners' medals between them, will have a home game against Southport if they beat Bradford Park Avenue in a replay.
Full draw (ties to be played on 24 October)
Gateshead v Worcester City
AFC Fylde v Barrow
Wrexham v Gainsborough Trinity
Northwich Victoria v Chorley
Harrogate Town v Grimsby Town
Barwell v AFC Rushden & Diamonds
Salford City or Bradford Park Ave v Southport
Sporting Khalsa v FC United or Buxton
Stalybridge Celtic v North Ferriby Utd
FC Halifax v Guiseley
Tranmere Rovers v Lincoln City
Stourbridge v Kidderminster Harriers
Macclesfield Town v Alfreton
Brackley v Bamber Bridge
Altrincham v Chester
Whitehawk v Poole Town
Maidenhead United v Woking
Basingstoke Town v Torquay United
Grays Athletic v Welling United
Boreham Wood v AFC Hornchurch
Wealdstone v Bognor Regis Town
Didcot Town v Brentwood Town
Eastbourne Borough v Dover Athletic
Chesham Utd v Enfield
Staines Town v Gloucester City
Aldershot Town v Sutton United
Bromley v Eastleigh
Margate v Forest Green Rovers
Braintree Town v Harlow Town
Havant and Waterlooville v Cheltenham Town
Chippenham Town v Maidstone United
St Albans City v Weston-super-Mare.
National League leaders Forest Green Rovers will play at Margate in the final qualifying round of the FA Cup.