Objectives: To examine race differences in knowledge of the Tuskegee study and the relationship between knowledge of the Tuskegee study and medical system mistrust. Methods: We conducted a telephone survey of 277 African-American and 101 white adults 18–93 years of age in Baltimore, MD. Participants responded to questions regarding mistrust of medical care, including a series of questions regarding the Tuskegee Study of Untreated Syphilis in the Negro Male (Tuskegee study). Results: Findings show no differences by race in knowledge of or about the Tuskegee study and that knowledge of the study was not a predictor of trust of medical care. However, we find significant race differences in medical care mistrust. Conclusions: Our results cast doubt on the proposition that the widely documented race difference in mistrust of medical care results from the Tuskegee study. Rather, race differences in mistrust likely stem from broader historical and personal experiences.