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Introduction: The aim of this study was to design a standard questionnaire facilitating the evaluation of the knowledge and attitude of medical students regarding radiation protection. Methods: At first, a 30-item questionnaire was prepared. The scale construction procedure was performed using content validity assessment. Considering objectives, some items were designed based on textbooks and the ideas of oral radiologists, medical physicists, and occupational medicine specialists as the expert panel. Content validity of the draft was determined by the panel. Results: Test-retest procedure was used to determine the reliability of the questionnaire by kappa statistic and Cronbach’s alpha coefficient. Experts evaluated the content validity as desirable. Kappa coefficient was more than 0.75 for almost all knowledge and attitude items. Cronbach’s alpha coefficients for basic knowledge, practical knowledge, and attitude domains were 0.793, 0.823, and 0.822, respectively. Conclusion: The designed questionnaire was confirmed as reliable considering Iranian cultural concepts.