Objective With advantages of different teaching approach, design and optimize a blended teaching model and learning resource based on study preference and rules of learning. Methods Analyze the study preference and study behaviors of students who have participated in the medical literature retrieval and application A in Fudan University in 2017. The 194 students include grade 2015 in clinical medicine major with 8-year-program and grade 2015 in Basic Medicine major with 5-year-program. The study behavior data were collected from the online platform, and the study preference data from questionnaire. Chisq analysis and ANOVA analysis were applied to analyze the data. Results The study preferences were significant different among four teaching methods (online autonomous learning, PBL, flipped classroom, traditional teaching) in different type of knowledge, such as introduction (χ²=36.01,P<0.001). For basic knowledge, students preferred the online autonomous learning; for comprehensive application and thinking method, students preferred traditional class teaching. There were significant differences in the discussion participation of students with different preferences in teaching form (χ²=17.87,P<0.001), but there was no significant difference in online self-test scores. In the form of learning resources, the PPT divided by knowledge points in PDF format is the most popular. Conclusions In blended teaching design, Teachers should not only focus on micro-video and flipped classroom, the learning characteristics of various kinds of knowledge should be fully considered. Comprehensive implementation of a variety of teaching forms should be applied in view of the students' learning preference. Teachers should give full play to the advantages of online autonomous learning, carefully design the PBL and flipped classroom to deepen the knowledge internalization, build the micro-grid and multi-dimensional learning resource system, and establish the mechanism of self-learning supervise.