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The 2000-2001 KNVB Cup (at the time called Amstel Cup) was the 83rd edition of the Dutch national football annual knockout tournament for the KNVB Cup. 86 teams contested, beginning on 8 August 2000 and ending at the final on 24 May 2001.
All 18 participants of the Eredivisie 2000-01: six teams entered in the round of 16 of the knock-out stage; one team in the first round of the knock-out stage and the rest in the group stage
The matches of the group stage were played between August 8 and September 13, 2000. 79 teams participated and 117 matches were played. 39 teams advanced to the next round.
The matches of the first round were played on September 20 and 21, 2000. RKC Waalwijk entered the tournament here, during the group stage they were still active in the Intertoto Cup.
The matches were played between January 24 and 27, 2001. The six Eredivisie teams that had been playing in European competitions after qualification last season, entered the tournament this round.