The 2000 Tuskegee Golden Tigers football team represented Tuskegee University as a member of the Southern Intercollegiate Athletic Conference (SIAC) during the 2000 NCAA Division II football season. Led by fifth-year head coach Rick Comegy, the Golden Tigers compiled an overall record of 12–0, with a conference record of 7–0, and finished as SIAC champion. At the conclusion of the season, the Golden Tigers were also recognized as black college national champion.
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