This study aims to determine the effect of Augmented Reality (AR) learning media using Assemblr Studio on students' technological literacy in Biology. The research method used was Quasi Experimental Design with a Post-test Only with Non-equivalent Group Design. The research sample consisted of two classes: class XI 4 as the experimental class using AR media with Assemblr Studio and class XI 1 as the control class using conventional methods in the form of PowerPoint subjects at MAN 2 Kota Tasikmalaya. The sampling technique used was purposive sampling. Data were collected through a posttest in the form of a technological literacy questionnaire using a Likert scale with 20 statements covering content, process, context, and attitude indicators. The hypothesis test results using the Independent Samples T-Test showed a significance value of 0.000 (<0.05), indicating a significant effect of using AR learning media with Assemblr Studio on students' technological literacy. In addition, the effect size (Cohen’s d) value of 1.14 is categorized as a high effect, demonstrating that the use of AR learning media with Assemblr Studio has a strong influence on technological literacy. This media has been proven to improve students' understanding, learning motivation, and technological literacy through a more realistic and engaging visual interactive learning experience, making it a potential alternative solution to enhance technological literacy in schools.