For this study, a scale was developed to measure the factors that may affect the gamification process in undergraduate education. The sequential exploratory mixed method was used to recruit the required data. Through the use of convenient sampling, 91 pre-service teachers participated in this study. Kahoot!, one of the online game platforms, was applied by the researchers to create a gamified learning environment, and a 10-week course plan was implemented. To collect the qualitative data, unstructured observations were undertaken each week. All the qualitative data were analysed via the content analysis method. Then, a scale initially including 26 items was developed based on the observation results and related literature. The quantitative data were gathered using this scale and exploratory factor analysis (EFA) was applied to examine the factor structure of the scale. The observation results were categorised under the following six themes: learning effect, expected outcome, competition, entertainment, engagement and intention. After required EFAs were implemented, three items were removed; thus, the final scale consisted of 23 items, and six factors that affect the gamification process in undergraduate education were revealed. The results concluded that the developed scale to measure the factors affecting the gamification process was remarkably valid and reliable.