The 2000–01 FA Cup qualifying rounds opened the 120th season of competition in England for 'The Football Association Challenge Cup' (FA Cup), the world's oldest association football single knockout competition. A total of 602 clubs were accepted for the competition, up 23 from the previous season’s 579.

This season saw invites extended to some teams from Level 10 in the English football pyramid. With the larger number of clubs entering the tournament from non-League teams (Levels 5 through 10), a new extra preliminary round was added to the tournament. As a result, the competition started with six rounds of preliminary (2) and qualifying (4) knockouts for these non-League teams.

The 32 winning teams from fourth qualifying round progressed to the First round proper, where League teams tiered at Levels 3 and 4 entered the competition.

Calendar Round Start date Leagues entering at this round New entries this round Winners from previous round Number of fixtures Extra preliminary round 19 August 2000 Levels 8-10 34 none 17 Preliminary round 2 September 2000 Level 7 387 17 202 First qualifying round 16 September 2000 none none 202 101 Second qualifying round 30 September 2000 Isthmian LeagueNorthern Premier LeagueSouthern Football League(Premier divisions) 67 101 84 Third qualifying round 14 October 2000 none none 84 42 Fourth qualifying round 28 October 2000 Football Conference 22 42 32

Extra preliminary round Matches in the first extra preliminary round since 1950–51 were played on the weekend beginning Saturday 19 August 2000. 34 clubs from Levels 8-10 of English football, entered at this stage of the competition, while other clubs from those levels get a bye to the next round.

Tie Home team (tier) Score Away team (tier)1Brigg Town (8)3–0Willington (9)2Eccleshill United (8)2–3Sheffield (8)3South Shields (9)2–1Garforth Town (8)4Tow Law Town (8)2–0St Helens Town (8)5Squires Gate (9)1–2West Auckland Town (8)6Guisborough Town (8)1–1Chadderton (9) Chadderton (9)4–1Guisborough Town (8)7Armthorpe Welfare (8)0–1Penrith (9)8Pelsall Villa (8)1–1Kidsgrove Athletic (8) Kidsgrove Athletic (8)1–3Pelsall Villa (8)

Tie Home team (tier) Score Away team (tier)9Holbeach United (8)1–0Halesowen Harriers (8)10Royston Town (10)0–5Newmarket Town (8)11Haringey Borough (10)4–1Concord Rangers (10)12Raunds Town (8)2–3Soham Town Rangers (8)13Buckingham Town (8)4–1Halstead Town (8)14Lowestoft Town (8)2–3Fakenham Town (8)15Yate Town (8)0–3Brislington (8)16Melksham Town (8)1–0Falmouth Town (10)17Wimborne Town (8)2–0Bridport (8)

Preliminary round Matches were played on weekend of Saturday 2 September 2000. A total of 404 clubs took part in this stage of the competition, including the 17 winners from the extra preliminary round, 300 clubs from Levels 8-10, who get a bye in the extra preliminary round and 87 clubs entering at this stage from the four divisions at Level 7 of English football (all, except Vauxhall Motors). The round featured 40 clubs from Level 10 in the competition, being the lowest ranked clubs in this round.

Tie Home team (tier) Score Away team (tier)1Louth United (9)1–5North Ferriby United (7)2Witton Albion (7)4–3Oldham Town (9)3Trafford (7)1–0Flixton (8)4Marske United (8)3–1South Shields (9)5Consett (8)2–1Hallam (8)6Yorkshire Amateur (9)5–0Parkgate (9)7Workington (7)0–1Farsley Celtic (7)8Bradford Park Avenue (7)1–0Brandon United (8)9Kennek Ryhope CA (9)0–5Salford City (8)10Whitley Bay (8)2–1Ashington (9)11Gretna (7)0–0Ramsbottom United (8)Ramsbottom United (8)3–2Gretna (7)12Glasshoughton Welfare (8)0–2West Auckland Town (8)13Billingham Synthonia (8)1–1Brigg Town (8)Brigg Town (8)1–0Billingham Synthonia (8)14Goole (8)3–0Jarrow Roofing BCA (8)15Castleton Gabriels (9)2–3Harrogate Railway Athletic (8)16Tow Law Town (8)2–2Newcastle Blue Star (8)Newcastle Blue Star (8)2–1Tow Law Town (8)17Chadderton (9)2–3Hebburn Town (8)18Prescot Cables (8)0–2Bedlington Terriers (8)19Maine Road (8)2–2Easington Colliery (8)Easington Colliery (8)3–2Maine Road (8)20Bacup Borough (9)0–4Harrogate Town (7)21Shotton Comrades (9)1–2Atherton Laburnum Rovers (9)22Clitheroe (8)4–1Shildon (9)23Mossley (8)4–1Hatfield Main (9)24Winsford United (7)1–1Selby Town (8)Selby Town (8)2–0Winsford United (7)25Durham City (8)4–0Thackley (8)26Skelmersdale United (8)1–1Guiseley (7)Guiseley (7)3–0Skelmersdale United (8)27Brodsworth Miners Welfare (8)2–2Blackpool Mechanics (9)Blackpool Mechanics (9)3–2Brodsworth Miners Welfare (8)28Horden Colliery Welfare (9)3–0Crook Town (8)29Darwen (9)2–2Penrith (9)Penrith (9)0–1Darwen (9)30Dunston Federation Brewery (8)2–0Curzon Ashton (8)31Great Harwood Town (8)2–0Northallerton Town (9)32Radcliffe Borough (7)3–1Rossington Main (9)33Denaby United (8)0–1Warrington Town (9)34Ossett Town (7)0–1Chorley (7)35Atherton Collieries (8)2–0Morpeth Town (8)36Evenwood Town (9)1–2Tadcaster Albion (9)37Woodley Sports (8)0–3Ashton United (7)38Sheffield (8)2–1Rossendale United (8)39Seaham Red Star (8)3–1Pontefract Collieries (9)40Peterlee Newtown (8)4–4Fleetwood Freeport (8)Fleetwood Freeport (8)3–0Peterlee Newtown (8)41Thornaby (9)4–0Esh Winning (9)42Abbey Hey (8)1–2Stocksbridge Park Steels (7)43Liversedge (8)1–1Chester-le-Street Town (8)Chester-le-Street Town (8)0–3Liversedge (8)44Billingham Town (8)2–0Ossett Albion (8)45Cheadle Town (8)0–1Bridlington Town (9)46Kendal Town (7)3–1Pickering Town (9)47Boston Town (8)2–3Leek CSOB (8)48Spalding United (7)4–0Glossop North End (8)49Bilston Town (7)3–0Sandwell Borough (8)50Staveley Miners Welfare (8)1–2Belper Town (7)51Gedling Town (9)5–1Boldmere St Michaels (8)52Oadby Town (8)2–1Willenhall Town (8)53Redditch United (7)3–2Arnold Town (8)54Newcastle Town (8)2–0Buxton (8)55Stamford (7)2–0Rushall Olympic (8)56Holbeach United (8)2–1Glapwell (8)57Atherstone United (7)2–1Pelsall Villa (8)58Bourne Town (8)1–2Shepshed Dynamo (7)59Corby Town (7)1–0Knypersley Victoria (8)60Cradley Town (8)1–2Paget Rangers (7)61Gresley Rovers (7)1–1Matlock Town (7)Matlock Town (7)4–1Gresley Rovers (7)62Racing Club Warwick (7)1–2Stourport Swifts (8)63West Midlands Police (8)1–1Bedworth United (7)Bedworth United (7)2–0West Midlands Police (8)64Eastwood Town (7)3–1Rocester (7)65Hinckley United (7)7–0Nantwich Town (8)66Stapenhill (8)1–0Bridgnorth Town (8)67Stratford Town (8)1–1Stafford Town (8)Stafford Town (8)Stratford Town (8)68Blakenall (7)0–1Oldbury United (8)69Borrowash Victoria (9)0–3Grantham Town (7)70Barwell (8)0–0Lincoln United (7)Lincoln United (7)2–4Barwell (8)71Blackstones (8)4–1Shifnal Town (8)72Congleton Town (7)0–0Wednesfield (8)Wednesfield (8)0–1Congleton Town (7)73Solihull Borough (7)2–1Mickleover Sports (9)74Stourbridge (8)0–0Rugby United (7)Rugby United (7)Stourbridge (8)75Chasetown (8)0–1Alfreton Town (8)76Sutton Coldfield Town (7)1–1Bromsgrove Rovers (7)Bromsgrove Rovers (7)3–2Sutton Coldfield Town (7)77Saffron Walden Town (10)1–2Histon (7)78Tiptree United (8)6–2Buckingham Town (8)79Flackwell Heath (9)0–2Bishop's Stortford (7)80Wroxham (8)0–3Ford United (7)81Felixstowe & Walton United (8)2–3Waltham Abbey (10)82Edgware Town (8)2–2Stowmarket Town (8)Stowmarket Town (8)1–0Edgware Town (8)83Chelmsford City (7)4–1Sawbridgeworth Town (10)84Northampton Spencer (8)0–0Southall (10)Southall (10)0–2Northampton Spencer (8)85Basildon United (10)w/o Yeading (7)Walkover for Yeading – Basildon United removed86Boreham Wood (7)3–0Great Wakering Rovers (8)87Bugbrooke St Michaels (8)0–3Newmarket Town (8)88Potters Bar Town (10)2–1Leighton Town (8)89Cogenhoe United (8)4–1Desborough Town (8)90Bowers United (10)1–1Bedford United (10)Bedford United (10)3–1Bowers United (10)91Chalfont St Peter (9)1–1Harwich & Parkeston (8)Harwich & Parkeston (8)Chalfont St Peter (9)92Ware (9)3–2Yaxley (8)93Romford (7)1–6Hemel Hempstead Town (8)94Wingate & Finchley (9)2–1Wivenhoe Town (8)95Woodbridge Town (8)3–0Marlow (8)96Baldock Town (7)2–0Ruislip Manor (10)97Uxbridge (7)6–1Holmer Green (10)98Barking (8)2–1Haringey Borough (10)99Wootton Blue Cross (8)2–2London Colney (10)London Colney (10)Wootton Blue Cross (8)