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Two major problems were identified in reviewing current research in Web-based instruction: 1) assessment of irrelevant learning outcome variables and 2) weak methodological design in conducting quantitative research. This quantitative study was designed to investigate whether different types of Web-browsing interface in Web-based instruction influence students' learning achievement of varied educational objectives. The studied interfaces were 1) browser scroll, 2) frame-based, and 3) pop-up-window interfaces. This study also investigated a potential interaction between the interface and students' prior knowledge, since prior knowledge had long been considered an important learning variable for acquiring new information. A total of 137 undergraduate students participated in the study. A statistical multivariate analysis of variance (MANOVA) was used to analyze collected data. Results showed significant main effects of independent variables in all criterion tests except for the treatment effect on a comprehension knowledge test.