The main aim of the study is to understand the change of the multicultural literacy in a multicultural education course. The multicultural education course was designed by the researcher depending on the analysis of literature in order to develop the pre-service teachers’ multicultural literacy. The quantitative and qualitative approaches were used to at the study. In the part of the quantitative approach, the researcher adopted the Scale of Pre-service Teacher’s Multicultural Literacy designed by Cai Chun Chun in 2006 and used t-test to compare the pre and the post score of the multicultural literacy. The result was found that multicultural education course could promote the pre-service teachers’ multicultural literacy including multicultural awareness, multicultural skills and the total scales. In the part of the qualitative approach, the researcher used the qualitative analyze system Nvivo7 to code and conclude. The result was found that multicultural education course could promote the pre-service teachers’ multicultural literacy, especially multicultural awareness.