 | 05.10.2020 15:00 | | Burnham | – | Whyteleafe | | 1:3 |
| Whyteleafe |
| 04.10.2020 15:00 | | Concord Rangers | – | Potters Bar Town | | 2:1 |
| Potters Bar |
| 04.10.2020 15:00 | | Havant & Waterlooville | – | Horsham | | 2:1 |
| Bognor Regis |
| 03.10.2020 15:00 | | Guiseley | – | Atherton Collieries | | 4:0 |
| Guiseley |
| 03.10.2020 15:00 | | Mossley AFC | – | Tadcaster Albion | | 1:1,
pen. 3:4 |
| Mossley |
| 03.10.2020 15:00 | | Chorley | – | Gateshead | | 2:1 |
| Chorley |
| 03.10.2020 15:00 | | Southport | – | Morpeth Town | | 2:1 |
| Southport |
| 03.10.2020 15:00 | | Stalybridge Celtic | – | Longridge Town | | 2:3 |
| Stalybridge |
| 03.10.2020 15:00 | | Whitley Bay | – | Blyth Spartans | | 2:4 |
| Whitley Bay |
| 03.10.2020 15:00 | | Farsley Celtic | – | Radcliffe | | 2:1 |
| Farsley |
| 03.10.2020 15:00 | | Runcorn Linnets | – | Marine | | 1:1,
pen. 3:4 |
| Runcorn |
| 03.10.2020 15:00 | | Ashton United | – | South Shields | | 0:4 |
| Ashton-under-Lyne |
| 03.10.2020 15:00 | | Curzon Ashton | – | United of Manchester | | 1:2 |
| Ashton-under-Lyne |
| 03.10.2020 15:00 | | Darlington | – | Prescot Cables | | 2:2,
pen. 5:4 |
| Darlington |
| 03.10.2020 15:00 | | Bradford Park Avenue | – | Spennymoor Town | | 1:3 |
| Bradford |
| 03.10.2020 15:00 | | Warrington Rylands 1906 | – | York City | | 0:1 |
| Warrington |
| 03.10.2020 15:00 | | Hyde | – | AFC Fylde | | 2:4 |
| Hyde |
| 03.10.2020 15:00 | | Marske United | – | Consett AFC | | 6:0 |
| Marske-by-the-Sea |
| 03.10.2020 15:00 | | Skelmersdale United | – | Lancaster City | | 2:1 |
| Skelmersdale |
| 03.10.2020 15:00 | | Boston United | – | AFC Mansfield | | 4:2 |
| Boston |
| 03.10.2020 15:00 | | Hednesford Town | – | Halesowen Town | | 0:0,
pen. 3:5 |
| Hednesford |
| 03.10.2020 15:00 | | Coalville Town | – | Alfreton Town | | 1:2 |
| Coalville |
| 03.10.2020 15:00 | | Tamworth | – | Evesham United | | 3:1 |
| Tamworth |
| 03.10.2020 15:00 | | Chasetown | – | A.F.C. Telford United | | 1:1,
pen. 4:5 |
| Chasetown |
| 03.10.2020 15:00 | | Alvechurch | – | Kidderminster Harriers | | 2:2,
pen. 4:2 |
| Alvechurch |
| 03.10.2020 15:00 | | Nantwich Town | – | Barwell | | 1:0 |
| Nantwich |
| 03.10.2020 15:00 | | Mickleover | – | Newark | | 4:1 |
| Mickleover |
| 03.10.2020 15:00 | | Buxton | – | Stafford Rangers | | 0:0,
pen. 2:4 |
| Buxton |
| 03.10.2020 15:00 | | Grantham Town | – | Matlock Town | | 0:1 |
| Grantham |
| 03.10.2020 15:00 | | Leamington | – | Banbury United | | 0:1 |
| Royal Leamington Spa |
| 03.10.2020 15:00 | | Worksop Town | – | Chester | | 2:2,
pen. 3:5 |
| Retford |
| 03.10.2020 15:00 | | Nuneaton Town | – | Stratford Town | | 2:1 |
| Nuneaton |
| 03.10.2020 15:00 | | Ilkeston | – | Hanley Town | | 4:1 |
| Ilkeston |
| 03.10.2020 15:00 | | AFC Dunstable | – | Hemel Hempstead Town | | 1:2 |
| Dunstable |
| 03.10.2020 15:00 | | Brackley Town | – | Billericay Town | | 2:2,
pen. 4:2 |
| Brackley |
| 03.10.2020 15:00 | | Maldon & Tiptree | – | Grays Athletic | | 2:2,
pen. 5:4 |
| Maldon |
| 03.10.2020 15:00 | | Canvey Island | – | Biggleswade | | 2:2,
pen. 4:3 |
| Canvey Island |
| 03.10.2020 15:00 | | Cheshunt | – | Cambridge City | | 1:2 |
| Cheshunt |
| 03.10.2020 15:00 | | Royston Town | – | Stamford | | 2:2,
pen. 4:2 |
| Royston |
| 03.10.2020 15:00 | | Barking | – | Kings Langley | | 2:2,
pen. 3:2 |
| London |
| 03.10.2020 15:00 | | Hashtag United | – | Braintree Town | | 1:1,
pen. 6:7 |
| Pitsea |
| 03.10.2020 15:00 | | St Albans City | – | Hitchin Town | | 5:0 |
| St Albans |
| 03.10.2020 15:00 | | Bishop's Stortford | – | Brentwood Town | | 1:0 |
| Bishop's Stortford |
| 03.10.2020 15:00 | | Walthamstow | – | AFC Hornchurch | | 0:2 |
| London |
| 03.10.2020 15:00 | | Leighton Town | – | Leiston | | 1:2 |
| Leighton Buzzard |
| 03.10.2020 15:00 | | Brantham Athletic | – | Aveley | | 0:3 |
| Brantham |
| 03.10.2020 15:00 | | Kettering Town | – | Chelmsford City | | 2:0 |
| Kettering |
| 03.10.2020 15:00 | | Peterborough Sports | – | Stansted | | 4:2 |
| Peterborough |
| 03.10.2020 15:00 | | Bury Town | – | Waltham Abbey | | 4:1 |
| Bury St Edmunds |
| 03.10.2020 15:00 | | Hayes & Yeading United | – | Bognor Regis Town | | 5:0 |
| London |
| 03.10.2020 15:00 | | Dartford | – | Slough Town | | 0:1 |
| Dartford |
| 03.10.2020 15:00 | | Farnborough | – | Tonbridge Angels | | 0:1 |
| Farnborough |
| 03.10.2020 15:00 | | Harrow Borough | – | Cray Valley Paper Mills | | 1:5 |
| London |
| 03.10.2020 15:00 | | Chipstead | – | East Grinstead Town | | 1:0 |
| Chipstead |
| 03.10.2020 15:00 | | Chichester City | – | Risborough Rangers | | 2:1 |
| Chichester |
| 03.10.2020 15:00 | | Corinthian-Casuals | – | Dulwich Hamlet | | 2:2,
pen. 1:3 |
| London |
| 03.10.2020 15:00 | | Haringey Borough | – | Chertsey Town | | 2:0 |
| London |
| 03.10.2020 15:00 | | Folkestone Invicta | – | Chatham Town | | 0:3 |
| Folkestone |
| 03.10.2020 15:00 | | Hendon | – | Maidstone United | | 0:1 |
| London |
| 03.10.2020 15:00 | | Hanwell Town | – | Hartley Wintney | | 3:5 |
| London |
| 03.10.2020 15:00 | | Moneyfields | – | Cray Wanderers | | 2:6 |
| Portsmouth |
| 03.10.2020 15:00 | | Ebbsfleet United | – | Hastings United | | 2:2,
pen. 4:1 |
| Northfleet |
| 03.10.2020 15:00 | | Sheppey United | – | Welling United | | 2:0 |
| Isle of Sheppey |
| 03.10.2020 15:00 | | Corinthian | – | Hampton & Richmond Borough | | 0:1 |
| Hartley Gay Dawn Farm |
| 03.10.2020 15:00 | | Bedfont Sports | – | Carshalton Athletic | | 2:0 |
| London |
| 03.10.2020 15:00 | | Dorking Wanderers | – | Eastbourne Borough | | 3:3,
pen. 3:4 |
| Dorking |
| 03.10.2020 15:00 | | Bracknell Town | – | Marlow | | 2:2,
pen. 4:3 |
| Bracknell Larges Lane |
| 03.10.2020 15:00 | | Metropolitan Police | – | Walton Casuals | | 1:2 |
| Molesey |
| 03.10.2020 15:00 | | Weston-super-Mare | – | Swindon Supermarine | | 2:2,
pen. 4:2 |
| Weston-super-Mare |
| 03.10.2020 15:00 | | Truro City | – | Hungerford Town | | 4:0 |
| Truro |
| 03.10.2020 15:00 | | Saltash United | – | Sholing | | 1:3 |
| Saltash |
| 03.10.2020 15:00 | | Frome Town | – | Larkhall Athletic | | 1:1,
pen. 3:4 |
| Frome |
| 03.10.2020 15:00 | | Christchurch | – | Gloucester City | | 1:1,
pen. 6:5 |
| Christchurch |
| 03.10.2020 15:00 | | Tiverton Town | – | Taunton Town | | 3:5 |
| Tiverton |
| 03.10.2020 15:00 | | Gosport Borough | – | Hereford | | 1:3 |
| Gosport |
| 03.10.2020 15:00 | | Bath City | – | Winchester City | | 3:2 |
| Bath |
| 03.10.2020 15:00 | | Wimborne Town | – | Melksham Town | | 0:0,
pen. 5:4 |
| Wimborne Minster |
| 03.10.2020 15:00 | | Kidlington | – | Bristol Manor Farm | | 1:1,
pen. 6:7 |
| Kidlington |
| 03.10.2020 15:00 | | Royal Wootton Bassett Town | – | Oxford City | | 1:2 |
| Wootton Bassett |
| 03.10.2020 15:00 | | Chippenham Town | – | Poole Town | | 2:2,
pen. 5:4 |
| Chippenham |
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