This study aims to research the effect of the interface background color choice on the emotion of university students. The study group consisted of 929 Turkish students studying at the vocational colleges of technical and social sciences in Çanakkale Onsekiz Mart University, Turkey. In this study, the descriptive method was used. A color effect form developed by the researcher was used. The validity of the tool was controlled by the faculty members in the Guidance and Psychological Counseling Department of Canakkale Onsekiz Mart University. The analysis of the findings showed that primary, secondary, and tertiary colors had positive impacts on the participants' perceptions except for gray color. In this negative perception, the boredom perception of the gray was affective. The colors having the most positive perceptions of the participants were white, blue, and red. In the positive perception of these colors, their perception of relaxing and encouraging was effective. In the colors' effect, there were significant differences according to the gender and academic education department. Red, black, white, gray, and purple colors' effect changed according to gender, while red, blue, and purple colors' effect changed according to the education department. As a result, this study, which is conducted with the participation of a large number and the usage of software, is expected to be a good reference on color perception and color design for software interface designers.