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Abstract Background This analysis assessed whether Blacks, Whites and Puerto-Rican (PR) Hispanics differed in their ability to identify the Tuskegee Syphilis Study (TSS) via open-ended questions following lead-in recognition and recall questions. Methods The Tuskegee Legacy Project (TLP) Questionnaire was administered via a Random-Digit Dial (RDD) telephone survey to a stratified random sample of Black, White and PR Hispanic adults in three U.S. cities. Results The TLP Questionnaire was administered to 1,162 adults (356 African-Americans, 313 PR Hispanics, and 493 non-Hispanic Whites) in San Juan, PR, Baltimore, MD and New York City, NY. Recall question data revealed: 1) that 89% or more of Blacks, Whites, and PR Hispanics were not able to name or definitely identify the Tuskegee Syphilis Study by giving study attributes; and, 2) that Blacks were the most likely to provide an open-ended answer that identified the Tuskegee Syphilis Study as compared to Whites and PR Hispanics (11.5% vs 6.3% vs 2.9%, respectively) (p ≤ 0.002). Even when probed by a recognition question, only a minority of each racial/ethnic group (37.1%, 26.9%, and 8.6%, for Blacks, Whites and PR Hispanics, respectively) was able to clearly identify the TSS (p < 0.001). Conclusions The two major implications of these findings for health disparity researchers are 1) that it is unlikely that detailed knowledge of the Tuskegee Syphilis Study has any current widespread influence on the willingness of minorities to participate in biomedical research, and 2) that caution should be applied before assuming that what community leaders 'know and are aware of' is equally 'well known' within their community constituencies.