This study aimed to examine the motivation of secondary school students toward context-based biology in terms of variables. Motivations were examined in terms of gender, grade level, school type, project experience, and biology laboratory experience. The research was planned according to the ‘Survey design.’ The sample consisted of 545 students studying at different grade levels (9th, 10th, 11th, and 12th grades) of secondary education institutions in Turkey. The study data were collected through the ‘Context-Based Biology Motivation Scale (CBBMS)’ developed by Gül (2019). The measurement tool consists of 34 items in a five-point Likert type, and a total of three sub-factors: association and effect, participation and competence, and enjoyment and satisfaction. Cronbach’s alpha reliability coefficient was determined (α = 0.943) for the entire measurement tool. For the sub-factors, it was α = 0.898 for association and effect, α = 0.914 for participation and competence, α = 0.846 and for enjoyment and satisfaction. For independent groups, the t-test, one-way analysis of variance (ANOVA), and Tukey test were used in the data analysis. According to the research results, secondary school students’ motivations toward context-based biology differed significantly according to gender, school type, biology laboratory experience, and grade level. On the contrary, motivations did not differ significantly according to their project experiences. The data obtained from the research will contribute to the literature in determining the factors affecting the motivation of secondary school students toward context-based biology.