| 14.09.2020 19:45 | | Basingstoke Town | | Chichester City | | 2:2, pen. 1:3 |
| Basingstoke |
| 13.09.2020 16:00 | | Hashtag United | | Felixstowe & Walton United | | 1:1, pen. 13:12 |
| Pitsea |
| 13.09.2020 15:00 | | Fawley | | Tadley Calleva | | 0:1 |
| Holbury |
| 12.09.2020 15:00 | | Whitley Bay | | Dunston UTS | | +:- |
| Whitley Bay |
| 12.09.2020 15:00 | | Penrith | | West Allotment Celtic | | 1:3 |
| Penrith |
| 12.09.2020 15:00 | | Kendal Town | | Bishop Auckland | | 0:5 |
| Kendal |
| 12.09.2020 15:00 | | West Auckland Town | | Ashington | | 3:3, pen. 3:0 |
| West Auckland |
| 12.09.2020 15:00 | | Billingham Town | | Stockton Town | | 1:2 |
| Stockton-on-Tees |
| 12.09.2020 15:00 | | Knaresborough Town | | Workington | | 1:3 |
| Knaresborough |
| 12.09.2020 15:00 | | Crook Town AFC | | Marske United | | 0:2 |
| Crook |
| 12.09.2020 15:00 | | Thornaby | | Sunderland RCA | | 0:2 |
| Thornaby-on-Tees |
| 12.09.2020 15:00 | | Frickley Athletic | | Newcastle Benfield | | 3:1 |
| South Elmsall |
| 12.09.2020 15:00 | | Hebburn Town | | Pontefract Collieries | | 2:2, pen. 4:5 |
| Hebburn |
| 12.09.2020 15:00 | | Guisborough Town | | Tadcaster Albion | | 1:2 |
| Guisborough |
| 12.09.2020 15:00 | | Consett AFC | | Ossett United | | 2:2, pen. 5:4 |
| Consett |
| 12.09.2020 15:00 | | Congleton Town | | Skelmersdale United | | 1:2 |
| Congleton |
| 12.09.2020 15:00 | | Glossop North End | | City of Liverpool | | 0:3 |
| Glossop |
| 12.09.2020 15:00 | | Ramsbottom United | | Irlam | | 4:1 |
| Ramsbottom |
| 12.09.2020 15:00 | | Ashton United | | Squires Gate | | 2:0 |
| Ashton-under-Lyne |
| 12.09.2020 15:00 | | Wythenshawe Amateurs | | Trafford | | 1:4 |
| Manchester |
| 12.09.2020 15:00 | | Runcorn Linnets | | Albion Sports | | 2:0 |
| Runcorn |
| 12.09.2020 15:00 | | Droylsden | | Litherland REMYCA | | -:+ |
| Droylsden |
| 12.09.2020 15:00 | | Silsden AFC | | Bootle | | 2:5 |
| Silsden |
| 12.09.2020 15:00 | | Mossley AFC | | St Helens Town | | 3:0 |
| Mossley |
| 12.09.2020 15:00 | | Northwich Victoria | | Charnock Richard | | 2:2, pen. 4:5 |
| Northwich |
| 12.09.2020 15:00 | | Colne | | Prescot Cables | | 0:2 |
| Colne |
| 12.09.2020 15:00 | | Runcorn Town | | Brighouse Town | | 1:1, pen. 5:4 |
| Runcorn |
| 12.09.2020 15:00 | | Warrington Rylands 1906 | | Clitheroe | | 1:0 |
| Warrington |
| 12.09.2020 15:00 | | Marine | | Barnoldswick Town | | 2:1 |
| Crosby |
| 12.09.2020 15:00 | | Widnes | | Longridge Town | | 2:3 |
| Widnes |
| 12.09.2020 15:00 | | Worcester City | | Walsall Wood | | 2:1 |
| Worcester |
| 12.09.2020 15:00 | | Black Country Rangers | | Matlock Town | | 0:1 |
| Tividale |
| 12.09.2020 15:00 | | Coventry Sphinx | | Coleshill Town | | 0:0, pen. 4:3 |
| Coventry |
| 12.09.2020 15:00 | | Sutton Coldfield Town | | Belper Town | | 0:1 |
| Sutton Coldfield |
| 12.09.2020 15:00 | | Kidsgrove Athletic | | Chasetown | | 0:1 |
| Kidsgrove |
| 12.09.2020 15:00 | | Leek Town | | Sporting Khalsa | | 2:1 |
| Leek |
| 12.09.2020 15:00 | | Westfields | | Whitchurch Alport | | 2:0 |
| Hereford |
| 12.09.2020 15:00 | | Racing Club Warwick | | Bedworth United | | 1:1, pen. 2:3 |
| Warwick |
| 12.09.2020 15:00 | | Market Drayton Town | | Tividale | | 0:1 |
| Market Drayton |
| 12.09.2020 15:00 | | Newcastle Town | | Halesowen Town | | 1:2 |
| Newcastle-under-Lyme |
| 12.09.2020 15:00 | | Evesham United | | Coventry United | | +:- |
| Evesham |
| 12.09.2020 15:00 | | Hanley Town | | Gresley Rovers | | 5:1 |
| Hanley |
| 12.09.2020 15:00 | | Shifnal Town | | AFC Bridgnorth | | 4:1 |
| Shifnal |
| 12.09.2020 15:00 | | Cleethorpes Town | | AFC Mansfield | | 0:1 |
| Grimsby |
| 12.09.2020 15:00 | | Boston Town | | Coalville Town | | 0:3 |
| Boston |
| 12.09.2020 15:00 | | Anstey Nomads | | Worksop Town | | 1:2 |
| Anstey |
| 12.09.2020 15:00 | | Holbeach United | | Sheffield | | 0:4 |
| Holbeach |
| 12.09.2020 15:00 | | Quorn | | Barton Town FC | | 2:0 |
| Quorn |
| 12.09.2020 15:00 | | Maltby Main | | Newark | | 0:4 |
| Maltby |
| 12.09.2020 15:00 | | West Bridgford | | Lincoln United | | +:- |
| West Bridgford Regatta Way Sports Ground |
| 12.09.2020 15:00 | | Ilkeston | | Shepshed Dynamo | | 3:0 |
| Ilkeston |
| 12.09.2020 15:00 | | Long Eaton United | | Bottesford Town | | 1:1, pen. 4:2 |
| Long Eaton |
| 12.09.2020 15:00 | | Oadby Town | | Carlton Town | | +:- |
| Oadby |
| 12.09.2020 15:00 | | Lutterworth Town | | Loughborough Dynamo | | 0:6 |
| Lutterworth |
| 12.09.2020 15:00 | | Wroxham | | Gorleston | | 3:1 |
| Wroxham |
| 12.09.2020 15:00 | | Biggleswade | | Histon | | 1:0 |
| Biggleswade |
| 12.09.2020 15:00 | | Cambridge City | | Biggleswade United | | 4:0 |
| Cambridge |
| 12.09.2020 15:00 | | Yaxley | | Stowmarket Town | | 1:2 |
| Yaxley |
| 12.09.2020 15:00 | | Mildenhall Town | | Corby Town | | 1:1, pen. 4:3 |
| Mildenhall |
| 12.09.2020 15:00 | | Ely City | | Eynesbury Rovers | | 3:1 |
| Ely |
| 12.09.2020 15:00 | | Stamford | | Diss Town | | 4:0 |
| Stamford |
| 12.09.2020 15:00 | | Thetford Town | | Potton United | | 0:2 |
| Blofield |
| 12.09.2020 15:00 | | Dereham Town | | Whitton United | | 3:2 |
| Dereham |
| 12.09.2020 15:00 | | Daventry Town | | Bedford Town | | 1:0 |
| Daventry |
| 12.09.2020 15:00 | | AFC Sudbury | | Harborough Town | | 4:2 |
| Sudbury |
| 12.09.2020 15:00 | | Haverhill Rovers | | Wisbech Town | | 4:3 |
| Haverhill |
| 12.09.2020 15:00 | | Cogenhoe United | | Bury Town | | 0:1 |
| Cogenhoe |
| 12.09.2020 15:00 | | Long Melford | | AFC Kempston Rovers | | 0:0, pen. 4:2 |
| Long Melford |
| 12.09.2020 15:00 | | Soham Town Rangers | | St Neots Town | | 3:2 |
| Soham |
| 12.09.2020 15:00 | | Royston Town | | Newmarket Town | | 6:0 |
| Royston |
| 12.09.2020 15:00 | | Great Wakering Rovers | | Brantham Athletic | | 0:1 |
| Great Wakering |
| 12.09.2020 15:00 | | Welwyn Garden City | | Saffron Walden Town | | 2:0 |
| Welwyn Garden City |
| 12.09.2020 15:00 | | Grays Athletic | | Witham Town | | 1:0 |
| Corringham |
| 12.09.2020 15:00 | | Coggeshall Town | | Tilbury | | 2:0 |
| Coggeshall |
| 12.09.2020 15:00 | | Walthamstow | | Cockfosters | | 1:1, pen. 3:2 |
| London |
| 12.09.2020 15:00 | | Sawbridgeworth Town | | St Margaretsbury | | 1:3 |
| Sawbridgeworth |
| 12.09.2020 15:00 | | FC Romania | | Brentwood Town | | 0:1 |
| Cheshunt |
| 12.09.2020 15:00 | | Barking | | Heybridge Swifts | | 2:0 |
| London |
| 12.09.2020 15:00 | | Clacton | | Hadley | | 1:1, pen. 10:9 |
| Clacton-on-Sea |
| 12.09.2020 15:00 | | Hertford Town | | Maldon & Tiptree | | 0:1 |
| Hertford |
| 12.09.2020 15:00 | | Bowers & Pitsea | | Barton Rovers | | 5:1 |
| Pitsea |
| 12.09.2020 15:00 | | Stansted | | Basildon United | | 2:0 |
| Stansted Mountfitchet |
| 12.09.2020 15:00 | | Harlow Town | | Sporting Bengal United | | 3:0 |
| Harlow |
| 12.09.2020 15:00 | | Harpenden Town | | Aveley | | 0:3 |
| Harpenden |
| 12.09.2020 15:00 | | Canvey Island | | Ware | | 2:0 |
| Canvey Island |
| 12.09.2020 15:00 | | Leiston | | Halstead Town | | 5:0 |
| Leiston |
| 12.09.2020 15:00 | | Waltham Abbey | | Woodford Town | | 1:0 |
| Waltham Abbey |
| 12.09.2020 15:00 | | Oxhey Jets | | Chalfont St Peter | | 0:1 |
| South Oxhey |
| 12.09.2020 15:00 | | Aylesbury United | | Long Crendon | | 3:0 |
| Aylesbury |
| 12.09.2020 15:00 | | Risborough Rangers | | Winslow United | | 2:1 |
| Princes Risborough |
| 12.09.2020 15:00 | | Flackwell Heath | | Cirencester Town | | 0:3 |
| Flackwell Heath |
| 12.09.2020 15:00 | | Didcot Town | | Royal Wootton Bassett Town | | 1:2 |
| Didcot |
| 12.09.2020 15:00 | | Holmer Green | | Highworth Town | | 0:2 |
| Holmer Green |
| 12.09.2020 15:00 | | Dunstable Town | | Easington Sports | | 1:1, pen. 7:6 |
| Dunstable |
| 12.09.2020 15:00 | | Kidlington | | Thame United | | 1:0 |
| Kidlington |
| 12.09.2020 15:00 | | Cribbs | | Berkhamsted | | 2:0 |
| Bristol |
| 12.09.2020 15:00 | | Harefield United | | Leighton Town | | 1:2 |
| London |
| 12.09.2020 15:00 | | Brimscombe & Thrupp | | Cinderford Town | | 1:2 |
| Brimscombe & Thrupp |
| 12.09.2020 15:00 | | Fairford Town | | Edgware Town | | 2:2, pen. 4:2 |
| Fairford |
| 12.09.2020 15:00 | | Northwood | | Slimbridge AFC | | 1:1, pen. 4:2 |
| London |
| 12.09.2020 15:00 | | Wantage Town | | Windsor | | 3:2 |
| Wantage |
| 12.09.2020 15:00 | | Marlow | | North Leigh | | 2:0 |
| Marlow |
| 12.09.2020 15:00 | | Thatcham Town | | AFC Dunstable | | 1:5 |
| Thatcham |
| 12.09.2020 15:00 | | Leverstock Green | | Burnham | | 1:4 |
| Leverstock Green |
| 12.09.2020 15:00 | | Tunbridge Wells | | Beckenham Town | | 1:1, pen. 4:1 |
| Royal Tunbridge Wells |
| 12.09.2020 15:00 | | Cray Valley Paper Mills | | VCD Athletic | | 6:0 |
| London |
| 12.09.2020 15:00 | | Hanwell Town | | Spelthorne Sports | | 4:1 |
| London |
| 12.09.2020 15:00 | | Sevenoaks Town | | CB Hounslow United | | 3:0 |
| Sevenoaks |
| 12.09.2020 15:00 | | Southall | | Ashford Town | | 2:1 |
| London |
| 12.09.2020 15:00 | | Chertsey Town | | Abbey Rangers | | 2:0 |
| Chertsey |
| 12.09.2020 15:00 | | Staines Town | | Guildford City | | 2:1 |
| Staines |
| 12.09.2020 15:00 | | East Grinstead Town | | Phoenix Sports | | 3:2 |
| East Grinstead |
| 12.09.2020 15:00 | | Hastings United | | Herne Bay | | 1:0 |
| Hastings |
| 12.09.2020 15:00 | | Ramsgate | | Chipstead | | 0:3 |
| Ramsgate |
| 12.09.2020 15:00 | | Sutton Common Rovers | | Broadfields United | | 4:0 |
| Sutton |
| 12.09.2020 15:00 | | Carshalton Athletic | | Whitstable Town | | 5:1 |
| Carshalton |
| 12.09.2020 15:00 | | Bedfont Sports | | Hassocks | | 3:1 |
| London |
| 12.09.2020 15:00 | | Deal Town | | Sittingbourne | | 4:1 |
| Deal |
| 12.09.2020 15:00 | | Newhaven | | Corinthian | | 1:2 |
| Newhaven |
| 12.09.2020 15:00 | | Erith & Belvedere | | Alfold | | 1:1, pen. 4:3 |
| London |
| 12.09.2020 15:00 | | Kingstonian | | Horley Town | | 4:1 |
| Kingston upon Thames |
| 12.09.2020 15:00 | | Westside | | Chatham Town | | 0:2 |
| London |
| 12.09.2020 15:00 | | Cobham | | Three Bridges | | 1:1, pen. 4:3 |
| Cobham |
| 12.09.2020 15:00 | | Whyteleafe | | Peacehaven & Telscombe | | 4:0 |
| Whyteleafe |
| 12.09.2020 15:00 | | Egham Town | | Crawley Down Gatwick | | 1:3 |
| Egham |
| 12.09.2020 15:00 | | Steyning Town | | Hanworth Villa | | 2:1 |
| Steyning |
| 12.09.2020 15:00 | | Broadbridge Heath | | Haywards Heath Town | | +:- |
| Broadbridge Heath |
| 12.09.2020 15:00 | | Sheppey United | | Uxbridge | | 4:1 |
| Isle of Sheppey |
| 12.09.2020 15:00 | | Hythe Town | | South Park | | 1:2 |
| Hythe |
| 12.09.2020 15:00 | | Tooting & Mitcham United | | Fisher | | 2:2, pen. 1:3 |
| London |
| 12.09.2020 15:00 | | Stansfeld | | Little Common | | 2:3 |
| London |
| 12.09.2020 15:00 | | Faversham Town | | Eastbourne Town | | 1:1, pen. 6:5 |
| Faversham |
| 12.09.2020 15:00 | | Ashford United | | Whitehawk | | 2:0 |
| Ashford |
| 12.09.2020 15:00 | | Burgess Hill Town | | Hollands & Blair | | +:- |
| Burgess Hill |
| 12.09.2020 15:00 | | Farnborough | | Lymington Town | | 0:0, pen. 4:3 |
| Farnborough |
| 12.09.2020 15:00 | | Hartley Wintney | | Hamworthy United | | 1:0 |
| Hartley Wintney |
| 12.09.2020 15:00 | | Bashley | | Christchurch | | 0:2 |
| Bashley |
| 12.09.2020 15:00 | | Westfield | | Frimley Green | | 2:2, pen. 3:5 |
| Woking |
| 12.09.2020 15:00 | | Moneyfields | | Camberley Town | | 4:2 |
| Portsmouth |
| 12.09.2020 15:00 | | Winchester City | | Corsham Town | | 2:0 |
| Winchester |
| 12.09.2020 15:00 | | Cowes Sports | | Hamble Club | | 1:0 |
| Cowes |
| 12.09.2020 15:00 | | Badshot Lea | | Bracknell Town | | 0:2 |
| Wrecclesham |
| 12.09.2020 15:00 | | Binfield | | AFC Totton | | 5:1 |
| Binfield |
| 12.09.2020 15:00 | | Bemerton Heath Harlequins | | Sholing | | 0:1 |
| Bemerton |
| 12.09.2020 15:00 | | Wimborne Town | | AFC Portchester | | 3:0 |
| Wimborne Minster |
| 12.09.2020 15:00 | | AFC Stoneham | | Pagham | | 3:0 |
| Eastleigh |
| 12.09.2020 15:00 | | Frome Town | | Bodmin Town | | 3:0 |
| Frome |
| 12.09.2020 15:00 | | Cadbury Heath | | Bristol Manor Farm | | 1:5 |
| Cadbury Heath |
| 12.09.2020 15:00 | | Exmouth Town | | Melksham Town | | 0:2 |
| Exmouth |
| 12.09.2020 15:00 | | Saltash United | | Paulton Rovers | | 1:0 |
| Saltash |
| 12.09.2020 15:00 | | Shepton Mallet | | Willand Rovers | | 1:0 |
| Shepton Mallet |
| 12.09.2020 15:00 | | Sherborne Town | | Clevedon Town | | 1:2 |
| Sherborne |
| 12.09.2020 15:00 | | Newton Abbot Spurs | | Larkhall Athletic | | 0:4 (0:2) |
| Newton Abbot |
| 12.09.2020 15:00 | | Bideford | | Wells City | | 3:0 |
| Bideford |
| 12.09.2020 15:00 | | Bridgwater Town | | Bitton | | 2:3 |
| Bridgwater |
| 12.09.2020 15:00 | | Barnstaple Town | | Helston Athletic | | 2:0 |
| Barnstaple |
| 12.09.2020 15:00 | | Tavistock | | Mangotsfield United | | 2:1 |
| Tavistock |
| 12.09.2020 12:30 | | Stocksbridge Park Steels | | Stalybridge Celtic | | 1:3 |
| Stocksbridge |
| 12.09.2020 11:30 | | Langney Wanderers | | Harrow Borough | | 1:1, pen. 2:3 |
| Eastbourne |
| 11.09.2020 19:45 | | New Salamis | | West Essex | | 5:1 |
| London |