The remaining Pitching In Northern Premier League Premier Division sides join the competition at this round, with a number of them drawn together.
In a reverse of the 2018/19 NPL play-off final, Warrington Town will host South Shields, while Scarborough Athletic face Ashton United, who were one of three Premier Division sides involved in Preliminary Round ties at the weekend.
There are a total of 12 ties that see NPL clubs go up against each other, including Pontefract Collieries travelling to FC United and Chasetown hosting Basford United.
The games will take place on Tuesday September 22. The winners receive £2,250 in prize money, with the losers pocketing £750.
If a game is drawn after 90 minutes, the winner will be determined by a penalty shoot-out in the absence of replays this season.
There are a total of 117 ties at this round of the competition.
The ties involving NPL clubs are below:
Sunderland RCA v Prescot Cables
Lancaster City v Runcorn Town
Warrington Town v South Shields
Marske United v Trafford
Mossley v Ramsbottom United
West Allotment Celtic v Hyde United
West Auckland Town v Runcorn Linnets
Whitley Bay v Witton Albion
Radcliffe v Workington
Scarborough Athletic v Ashton United
Frickley Athletic v Marine
Whitby Town v Warrington Rylands
City of Liverpool v Morpeth Town
Atherton Collieries v Bamber Bridge
Stalybridge Celtic v Bishop Auckland
FC United v Pontefract Collieries
Tadcaster Albion v Litherland REMYCA
Worcester City v Stafford Rangers
Quorn v Matlock Town
Leek Town v Mickleover
Grantham Town v Rushall Olympic
Nuneaton Borough v Loughborough Dynamo
Westfields v Worksop Town
Coventry Sphinx v Ilkeston Town
Banbury United v Carlton Town
Chasetown v Basford United
Tividale v Nantwich Town
AFC Mansfield v Gainsborough Trinity
Coalville Town v Sheffield
Buxton v Belper Town
Stamford v AFC Sudbury