This study aims to analyze the effectiveness of bioinformatics-based molecular genetics learning in improving students’ 21st-century competencies. A quantitative approach with a pre-experimental one group pretest-posttest design was employed. The participants consisted of students from one class selected through total sampling. The research procedure included administering a pretest, implementing bioinformatics-based learning using NCBI BLAST supported by student worksheets, and conducting a posttest. Data were analyzed using descriptive statistics, N-gain, and a paired sample t-test. The results showed that the mean pretest score of 72.30 increased to 87.95 in the posttest. The N-gain value was 0.56, categorized as moderate. The paired sample t-test indicated a significant difference between pretest and posttest scores (p<0.05). These findings demonstrate that bioinformatics-based learning effectively enhances students’ 21st-century competencies, including critical thinking, problem-solving, digital literacy, and collaboration. Therefore, integrating bioinformatics into molecular genetics learning can serve as an innovative approach to improve learning quality in higher education.
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