| 13 Nov 21 | r01  | Marske UnitedMarske United | 3 - 2 | FC United of ManchesterFC United of |
| 13 Nov 21 | r01 | Nantwich TownNantwich Town | 2 - 1 | Grantham TownGrantham Town |
| 13 Nov 21 | r01 | ColneColne | 1 - 1 | TamworthTamworth |
| 13 Nov 21 | r01 | Whitby TownWhitby Town | 5 - 0 | MossleyMossley |
| 13 Nov 21 | r01 | GainsboroughGainsborough | 1 - 2 | Lancaster CityLancaster Cit |
| 13 Nov 21 | r01 | Brentwood TownBrentwood Tow | 4 - 1 | Staines TownStaines Town |
| 13 Nov 21 | r01 | Needham MarketNeedham Marke | 3 - 2 | MargateMargate |
| 13 Nov 21 | r01 | Hayes & Yeading UnitedHayes & Yeadi | 0 - 3 | WorthingWorthing |
| 13 Nov 21 | r01 | MarineMarine | 2 - 2 | Dunston UTSDunston UTS |
| 13 Nov 21 | r01 | Folkestone InvictaFolkestone In | 2 - 1 | Faversham TownFaversham Tow |
| 13 Nov 21 | r01 | FarnboroughFarnborough | 1 - 0 | BanburyBanbury |
| 13 Nov 21 | r01 | Weston-super-Mare FCWeston-super- | 1 - 2 | Wimborne TownWimborne Town |
| 13 Nov 21 | r01 | RadcliffeRadcliffe | 2 - 0 | Nuneaton TownNuneaton Town |
| 13 Nov 21 | r01 | Rushall OlympicRushall Olymp | 0 - 4 | MatlockMatlock |
| 13 Nov 21 | r01 | AshtonAshton | 5 - 1 | Sutton Coldfield TownSutton Coldfi |
| 13 Nov 21 | r01 | Warrington TownWarrington To | 2 - 0 | ChasetownChasetown |
| 13 Nov 21 | r01 | Bromsgrove SportingBromsgrove Sp | 0 - 0 | Morpeth TownMorpeth Town |
| 13 Nov 21 | r01 | Carlton TownCarlton Town | 1 - 2 | StourbridgeStourbridge |
| 13 Nov 21 | r01 | BootleBootle | 3 - 3 | StalybridgeStalybridge |
| 13 Nov 21 | r01 | LiversedgeLiversedge | 3 - 2 | Basford Utd.Basford Utd. |
| 13 Nov 21 | r01 | Walton CasualsWalton Casual | 2 - 6 | UxbridgeUxbridge |
| 13 Nov 21 | r01 | CanveyCanvey | 6 - 0 | YaxleyYaxley |
| 13 Nov 21 | r01 | Bishops StortfordBishops Stort | 2 - 1 | ChipsteadChipstead |
| 13 Nov 21 | r01 | Royston TownRoyston Town | 1 - 1 | KingstonianKingstonian |
| 13 Nov 21 | r01 | Welwyn Garden CityWelwyn Garden | 2 - 1 | Carshalton AthleticCarshalton At |
| 13 Nov 21 | r01 | CheshuntCheshunt | 1 - 0 | BerkhamstedBerkhamsted |
| 13 Nov 21 | r01 | Bedford TownBedford Town | - | Potters Bar TownPotters Bar T |
| 13 Nov 21 | r01 | LeistonLeiston | 3 - 0 | Harrow BoroughHarrow Boroug |
| 13 Nov 21 | r01 | Biggleswade TownBiggleswade T | 1 - 2 | Enfield TownEnfield Town |
| 13 Nov 21 | r01 | Felixstowe & Walton UniteFelixstowe & | 2 - 1 | Peterborough SportsPeterborough |
| 13 Nov 21 | r01 | Hanwell TownHanwell Town | 1 - 0 | Wingate & FinchleyWingate & Fin |
| 13 Nov 21 | r01 | Cray WanderersCray Wanderer | 2 - 1 | Bedfont SportsBedfont Sport |
| 13 Nov 21 | r01 | MarlowMarlow | 2 - 2 | Larkhall AthleticLarkhall Athl |
| 13 Nov 21 | r01 | Hartley WintneyHartley Wintn | 3 - 0 | Willand RoversWilland Rover |
| 13 Nov 21 | r01 | BinfieldBinfield | 3 - 0 | Cinderford TownCinderford To |
| 13 Nov 21 | r01 | AFC TottonAFC Totton | 2 - 2 | Frome TownFrome Town |
| 13 Nov 21 | r01 | Plymouth ParkwayPlymouth Park | 4 - 0 | SalisburySalisbury |
| 13 Nov 21 | r01 | Bracknell TownBracknell Tow | 1 - 3 | Tiverton TownTiverton Town |
| 14 Nov 21 | r01 | Truro CityTruro City | 1 - 1 | Bognor Regis TownBognor Regis |
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