This study aimed to analyze the effect of practicum-based learning on students’ learning interest, learning outcomes, and both variables simultaneously in biotechnology learning among twelfth-grade students at MA Tarbiyatul Mubtadiin. The study was motivated by teacher-centered instruction that limited students’ active participation. A quantitative quasi-experimental approach with a Nonequivalent Control Group Design was employed. The sample consisted of 44 students selected through purposive sampling, comprising 22 students in the experimental group and 22 students in the control group. Data were collected using a learning-interest questionnaire and an achievement test. Instruments were examined using Aiken’s V, Pearson Product Moment, and Cronbach’s Alpha. Data analysis included N-Gain, Shapiro–Wilk, Levene’s Test, Independent Samples t-Test, coefficient of determination, and MANOVA using SPSS version 25. The experimental group’s learning interest increased from 78.41% to 87.80%, while the control group increased from 78.86% to 87.65%. Learning outcomes increased from 67.50 to 88.45 in the experimental group and from 64.27 to 78.77 in the control group. The t-test produced Sig. 0.000 and t = 5.653, while MANOVA produced Wilks’ Lambda = 0.563 with Sig. 0.000. Practicum-based learning significantly affected learning interest, learning outcomes, and both variables simultaneously.
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