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This study aimed to determine the assessment of pre-service biology teachers on student worksheets based on Scientific Literacy. This research was conducted at the Biology EducationDepartment, Sultan AgengTirtayasaUniversity. The method used was a Quasi-experimental. The research design used was one-shot case study. The subjects in this study consisted of 30 pre-service biology teachers. The instrument used was assessment sheets for student worksheets based on scientific literacy. The results of this study indicate the average assessment of pre-service biology teachers on student worksheets based on Scientific Literacy is 3.8 (valid category). Thus it can be concluded that student worksheets based on Scientific Literacy are suitable to be used in the research process and learning process.