Biodiversity material is close to the lives of students. Hence the possibility of students having their own experience will be their initial knowledge on the material. In the Curriculum Merdeka, biodiversity material is taught in the odd semester of class X SMA. Therefore, to determine the preconceptions of students, a three tier test is given at the grade IX level of junior high school, specifically students who have not received the material on biodiversity. The purpose of this study was to develop a three-tier test that was used to determine the preconceptions of students. This research uses research and development method with 4D design which consists of define, design, develop and dissemination stages. The research sample used included 220 junior high school students determined by purposive sampling technique. There are 18 out of 36 items which are stated to have good validity and difficulty level. The developed three tier test was declared reliable with an average high category in the first trial of 5,201, the second trial of 3,853 and the third trial of 1,040. The quantitative and descriptive results of the use of a valid three tier test were found that the instrument could be used to measure students’ preconceptions on biodiversity materials. The results of this study can be used as a reference for teacher to develop appropriate teaching materials and learning strategies for teaching biodiversity materials.

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