 | 23.09.2020 19:45 | | Worcester City | – | Stafford Rangers | | 2:3 |
| Worcester |
| 23.09.2020 19:45 | | Grays Athletic | – | Potton United | | 3:1 |
| Corringham |
| 23.09.2020 19:45 | | Sutton Common Rovers | – | Metropolitan Police | | 1:3 |
| Sutton |
| 23.09.2020 19:45 | | Wingate & Finchley | – | Folkestone Invicta | | 1:4 |
| London |
| 23.09.2020 19:45 | | Cray Wanderers | – | Fisher | | 3:1 |
| London |
| 23.09.2020 19:45 | | Cray Valley Paper Mills | – | Burgess Hill Town | | 3:1 |
| London |
| 23.09.2020 19:45 | | Tavistock | – | Gosport Borough | | 2:2,
pen. 3:5 |
| Tavistock |
| 23.09.2020 19:45 | | Kidlington | – | Salisbury | | 1:1,
pen. 4:2 |
| Kidlington |
| 23.09.2020 19:45 | | Larkhall Athletic | – | Bitton | | 3:2 |
| Bath |
| 23.09.2020 19:45 | | Tiverton Town | – | Bideford | | 2:0 |
| Tiverton |
| 23.09.2020 19:45 | | Highworth Town | – | Melksham Town | | 1:1,
pen. 4:5 |
| Highworth |
| 22.09.2020 20:00 | | Skelmersdale United | – | Bootle | | 2:1 |
| Skelmersdale |
| 22.09.2020 20:00 | | Lowestoft Town | – | Aveley | | 2:3 |
| Lowestoft |
| 22.09.2020 19:45 | | Sunderland RCA | – | Prescot Cables | | 0:4 |
| Sunderland |
| 22.09.2020 19:45 | | Lancaster City | – | Runcorn Town | | 1:0 |
| Lancaster |
| 22.09.2020 19:45 | | Warrington Town | – | South Shields | | 0:1 |
| Warrington |
| 22.09.2020 19:45 | | Marske United | – | Trafford | | 1:0 |
| Marske-by-the-Sea |
| 22.09.2020 19:45 | | Mossley AFC | – | Ramsbottom United | | 2:1 |
| Mossley |
| 22.09.2020 19:45 | | West Allotment Celtic | – | Hyde | | 0:5 |
| Longbenton |
| 22.09.2020 19:45 | | West Auckland Town | – | Runcorn Linnets | | 0:1 |
| West Auckland |
| 22.09.2020 19:45 | | Whitley Bay | – | Witton Albion | | 3:2 |
| Whitley Bay |
| 22.09.2020 19:45 | | Radcliffe | – | Workington | | 5:3 |
| Radcliffe |
| 22.09.2020 19:45 | | Scarborough Athletic | – | Ashton United | | 0:2 |
| Scarborough |
| 22.09.2020 19:45 | | Frickley Athletic | – | Marine | | 0:1 |
| South Elmsall |
| 22.09.2020 19:45 | | Whitby Town | – | Warrington Rylands 1906 | | 1:2 |
| Whitby |
| 22.09.2020 19:45 | | City of Liverpool | – | Morpeth Town | | 0:3 |
| Liverpool |
| 22.09.2020 19:45 | | Atherton Collieries | – | Bamber Bridge | | +:- |
| Atherton |
| 22.09.2020 19:45 | | Longridge Town | – | Charnock Richard | | 2:0 |
| Longridge |
| 22.09.2020 19:45 | | Stalybridge Celtic | – | Bishop Auckland | | 3:0 |
| Stalybridge |
| 22.09.2020 19:45 | | Consett AFC | – | Stockton Town | | 1:0 |
| Consett |
| 22.09.2020 19:45 | | United of Manchester | – | Pontefract Collieries | | 6:2 |
| Bury |
| 22.09.2020 19:45 | | Tadcaster Albion | – | Litherland REMYCA | | 7:2 |
| Tadcaster |
| 22.09.2020 19:45 | | Hednesford Town | – | Long Eaton United | | 3:2 |
| Hednesford |
| 22.09.2020 19:45 | | Tamworth | – | Stourbridge | | 3:3,
pen. 5:4 |
| Tamworth |
| 22.09.2020 19:45 | | Quorn | – | Matlock Town | | 0:2 |
| Quorn |
| 22.09.2020 19:45 | | Leek Town | – | Mickleover | | 1:2 |
| Leek |
| 22.09.2020 19:45 | | Grantham Town | – | Rushall Olympic | | 2:2,
pen. 3:1 |
| Grantham |
| 22.09.2020 19:45 | | Nuneaton Town | – | Loughborough Dynamo | | 2:1 |
| Nuneaton |
| 22.09.2020 19:45 | | Westfields | – | Worksop Town | | 1:3 |
| Hereford |
| 22.09.2020 19:45 | | West Bridgford | – | Halesowen Town | | 0:1 |
| West Bridgford Regatta Way Sports Ground |
| 22.09.2020 19:45 | | Coventry Sphinx | – | Ilkeston | | 0:2 |
| Coventry |
| 22.09.2020 19:45 | | Banbury United | – | Carlton Town | | 2:0 |
| Banbury |
| 22.09.2020 19:45 | | Chasetown | – | Basford United | | 2:1 |
| Chasetown |
| 22.09.2020 19:45 | | Tividale | – | Nantwich Town | | 2:4 |
| Tividale |
| 22.09.2020 19:45 | | Barwell | – | Bedworth United | | 3:1 |
| Barwell |
| 22.09.2020 19:45 | | AFC Mansfield | – | Gainsborough Trinity | | 3:0 |
| Clipstone The Forest Town Stadium |
| 22.09.2020 19:45 | | Coalville Town | – | Sheffield | | 2:0 |
| Coalville |
| 22.09.2020 19:45 | | Buxton | – | Belper Town | | 7:0 |
| Buxton |
| 22.09.2020 19:45 | | Daventry Town | – | Evesham United | | 0:2 |
| Daventry |
| 22.09.2020 19:45 | | AFC Rushden & Diamonds | – | Newark | | 0:5 |
| Wellingborough |
| 22.09.2020 19:45 | | Shifnal Town | – | Alvechurch | | 0:2 |
| Shifnal |
| 22.09.2020 19:45 | | Bromsgrove Sporting | – | Stratford Town | | 1:2 |
| Bromsgrove |
| 22.09.2020 19:45 | | Hanley Town | – | Redditch United | | 3:2 |
| Hanley |
| 22.09.2020 19:45 | | Ely City | – | Biggleswade | | 1:2 |
| Ely |
| 22.09.2020 19:45 | | Haverhill Rovers | – | Maldon & Tiptree | | 0:3 |
| Haverhill |
| 22.09.2020 19:45 | | Walthamstow | – | St Margaretsbury | | 0:0,
pen. 4:3 |
| London |
| 22.09.2020 19:45 | | Bury Town | – | Brightlingsea Regent | | 2:1 |
| Bury St Edmunds |
| 22.09.2020 19:45 | | Dereham Town | – | Canvey Island | | 0:2 |
| Dereham |
| 22.09.2020 19:45 | | Kings Langley | – | Clacton | | 1:1,
pen. 4:2 |
| Kings Langley |
| 22.09.2020 19:45 | | Stamford | – | AFC Sudbury | | 4:0 |
| Stamford |
| 22.09.2020 19:45 | | Bowers & Pitsea | – | AFC Hornchurch | | 0:3 |
| Pitsea |
| 22.09.2020 19:45 | | Long Melford | – | Cheshunt | | 1:3 |
| Long Melford |
| 22.09.2020 19:45 | | Leiston | – | Biggleswade Town | | 5:1 |
| Leiston |
| 22.09.2020 19:45 | | Cambridge City | – | Stowmarket Town | | 1:1,
pen. 4:2 |
| Cambridge |
| 22.09.2020 19:45 | | Peterborough Sports | – | Enfield Town | | 2:2,
pen. 5:4 |
| Peterborough |
| 22.09.2020 19:45 | | Potters Bar Town | – | East Thurrock United | | 1:0 |
| Potters Bar |
| 22.09.2020 19:45 | | Harlow Town | – | Waltham Abbey | | 0:1 |
| Harlow |
| 22.09.2020 19:45 | | Royston Town | – | Wroxham | | 2:0 |
| Royston |
| 22.09.2020 19:45 | | Leighton Town | – | Mildenhall Town | | 4:0 |
| Leighton Buzzard |
| 22.09.2020 19:45 | | Welwyn Garden City | – | Bishop's Stortford | | 1:1,
pen. 3:4 |
| Welwyn Garden City |
| 22.09.2020 19:45 | | Coggeshall Town | – | Stansted | | 0:1 |
| Coggeshall |
| 22.09.2020 19:45 | | Barking | – | Dunstable Town | | 6:1 |
| London |
| 22.09.2020 19:45 | | Brantham Athletic | – | St Ives Town | | 1:0 |
| Brantham |
| 22.09.2020 19:45 | | Burnham | – | Northwood | | 1:0 |
| Burnham |
| 22.09.2020 19:45 | | Horsham | – | Kingstonian | | 2:1 |
| Horsham |
| 22.09.2020 19:45 | | Corinthian | – | Sevenoaks Town | | 3:1 |
| Hartley Gay Dawn Farm |
| 22.09.2020 19:45 | | Whyteleafe | – | Binfield | | 2:0 |
| Whyteleafe |
| 22.09.2020 19:45 | | Chipstead | – | Deal Town | | 1:1,
pen. 4:2 |
| Chipstead |
| 22.09.2020 19:45 | | Staines Town | – | Walton Casuals | | 1:2 |
| Staines |
| 22.09.2020 19:45 | | Frimley Green | – | Marlow | | 1:1,
pen. 1:4 |
| Frimley Green |
| 22.09.2020 19:45 | | Little Common | – | Corinthian-Casuals | | 0:3 |
| Bexhill-on-Sea |
| 22.09.2020 19:45 | | Haringey Borough | – | Tunbridge Wells | | 5:1 |
| London |
| 22.09.2020 19:45 | | Haywards Heath Town | – | Hanwell Town | | 0:1 |
| Haywards Heath |
| 22.09.2020 19:45 | | Cobham | – | Risborough Rangers | | 1:3 |
| Cobham |
| 22.09.2020 19:45 | | Ashford United | – | Bracknell Town | | 1:4 |
| Ashford |
| 22.09.2020 19:45 | | Bedfont Sports | – | Lewes | | 3:1 |
| London |
| 22.09.2020 19:45 | | Hastings United | – | Chesham United | | 0:0,
pen. 6:5 |
| Hastings |
| 22.09.2020 19:45 | | Steyning Town | – | Sheppey United | | 0:5 |
| Steyning |
| 22.09.2020 19:45 | | South Park | – | Bognor Regis Town | | 2:2,
pen. 2:4 |
| Reigate |
| 22.09.2020 19:45 | | Merstham | – | AFC Dunstable | | 2:2,
pen. 6:7 |
| Merstham |
| 22.09.2020 19:45 | | Chertsey Town | – | Leatherhead | | 0:0,
pen. 5:4 |
| Chertsey |
| 22.09.2020 19:45 | | Chatham Town | – | Southall | | 3:2 |
| Chatham |
| 22.09.2020 19:45 | | Chalfont St Peter | – | Farnborough | | 2:3 |
| Chalfont St Peter |
| 22.09.2020 19:45 | | Margate | – | Hayes & Yeading United | | 1:2 |
| Margate |
| 22.09.2020 19:45 | | Fairford Town | – | Sholing | | 1:2 |
| Fairford |
| 22.09.2020 19:45 | | Cinderford Town | – | Royal Wootton Bassett Town | | 2:2,
pen. 5:6 |
| Cinderford |
| 22.09.2020 19:45 | | Chichester City | – | Cribbs | | 3:1 |
| Chichester |
| 22.09.2020 19:45 | | Aylesbury United | – | Moneyfields | | 2:2,
pen. 3:4 |
| Aylesbury |
| 22.09.2020 19:45 | | Taunton Town | – | Wantage Town | | 5:0 |
| Taunton |
| 22.09.2020 19:45 | | Winchester City | – | Clevedon Town | | 3:2 |
| Winchester |
| 22.09.2020 19:45 | | Tadley Calleva | – | Truro City | | 0:1 |
| Tadley |
| 22.09.2020 19:45 | | Barnstaple Town | – | Wimborne Town | | 2:3 |
| Barnstaple |
| 22.09.2020 19:45 | | Yate Town | – | Bristol Manor Farm | | 1:2 |
| Yate |
| 22.09.2020 19:45 | | Merthyr Town | – | Poole Town | | -:+ |
| Merthyr Tydfil |
| 22.09.2020 19:45 | | Frome Town | – | AFC Stoneham | | 4:1 |
| Frome |
| 22.09.2020 19:45 | | Christchurch | – | Dorchester Town | | 2:1 |
| Christchurch |
| 22.09.2020 19:45 | | Swindon Supermarine | – | Shepton Mallet | | 3:0 |
| Swindon |
| 22.09.2020 19:45 | | Saltash United | – | Cirencester Town | | 3:1 |
| Saltash |
| 22.09.2020 19:30 | | Hartley Wintney | – | Erith & Belvedere | | 5:0 |
| Hartley Wintney |
| 22.09.2020 19:30 | | Crawley Down Gatwick | – | Hendon | | 1:2 |
| Crawley Down The Haven Sportsfield |
| 22.09.2020 19:30 | | East Grinstead Town | – | Worthing | | 3:3,
pen. 4:2 |
| East Grinstead |
| 22.09.2020 19:30 | | Cowes Sports | – | Weston-super-Mare | | 0:5 |
| Cowes |
| 21.09.2020 19:45 | | Hitchin Town | – | Needham Market | | 3:0 |
| Hitchin |
| 21.09.2020 19:45 | | Hashtag United | – | Soham Town Rangers | | 1:1,
pen. 4:2 |
| Pitsea |
| 21.09.2020 19:45 | | New Salamis | – | Brentwood Town | | 1:2 |
| London |
| 21.09.2020 19:45 | | Beaconsfield Town | – | Harrow Borough | | 0:2 |
| Beaconsfield |
| 21.09.2020 19:45 | | Carshalton Athletic | – | Faversham Town | | 5:0 |
| Carshalton |
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