A total of 31 National League North and South sides made it through the previous round on Saturday and have discovered which sides stand in their way next midweek.
Ties will all be settled on the day with no extra time played, any match level after 90 minutes will go straight to penalties.
Standout ties include Chorley facing York City and Bath City heading to Slough Town.
Winners will receive £5,625 from the prize fund, with defeated sides taking away £1,875 from the round.
Ties including National League representatives:
Southport vs South Shields
Chester vs Spennymoor Town
Marske United vs Blyth Spartans
Farsley Celtic vs AFC Fylde
Darlington vs Tadcaster Albion
Chorley vs York City
Guiseley vs Matlock Town
FC United of Manchester vs Alfreton Town
St Albans City vs Mickleover
Braintree Town vs Maldon & Tiptree
Stafford Rangers vs Hereford
Leiston vs AFC Telford United
Brackley Town vs Kettering Town
Oxford City vs Tamworth
Boston United vs Hemel Hempstead Town
Havant & Waterlooville vs Chatham Town
Eastbourne Borough vs Sheppey United
Slough Town vs Bath City
Christchurch vs Dulwich Hamlet
Wimborne Town vs Maidstone United
Chichester City vs Tonbridge Angels
Hampton & Richmond Borough vs Hornchurch
Burnham or Whyteleafe vs Concord Rangers
Ebbsfleet United vs Chippenham Town