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Background  The inverted classroom approach is characterized by a primary self-study phase for students followed by an on-site, face-to-face teaching phase that is used to deepen the prior acquired knowledge. Obviously, this teaching approach relies on the students preparing before the on-site phase, which in turn requires optimized preparatory material as well as defined working instructions. The major aim of this study, therefore, was to investigate the effect of different preparatory materials and working instructions for the self-study phase of an e-learning-based inverted classroom on the knowledge gained by medical students in biochemistry. Furthermore, we analyzed whether collaborative dyadic learning during the self-study phase is more effective than individual learning with respect to knowledge gain.    Methods  The study was performed in a biochemistry seminar for second semester medical students at Ulm University in Germany. This seminar was held using an e-learning-based inverted classroom. A total of 196 students were divided into three homogeneous study groups that differed in terms of the working material and instructions provided for the self-study phase. Knowledge gain was measured by formative tests at the beginning of the on-site phases. Questionnaires were also handed out asking about motivation, interest and learning time in the self-study phases.    Results  Students who were told to prepare in collaborating dyads during the self-study phase performed better in formative tests taken at the beginning of on-site phases than learners who were told to prepare individually. The study material that was provided was of minor importance for the differences in formative testing since almost all students prepared for the on-site phases. With the dyadic learning approach, both students benefited from this collaboration, characterized by a higher motivation and interest in the topic, as well as a longer time spent on task.    Conclusion  Our study provides strong evidence that the study material, but more importantly the instructions provided for the self-study phase, affect students` knowledge gain in an e-learning-based inverted classroom. The instructed collaboratively working group was the most successful.    Electronic supplementary material  The online version of this article (10.1186/s12909-019-1497-y) contains supplementary material, which is available to authorized users.

Investigating the self-study phase of an inverted biochemistry classroom – collaborative dyadic learning makes the difference