Students negative emotions, beliefs, and behaviours toward the study of mathematics have tremendously contributed to their poor performance in the subject and it is caused by factors emanating from home, socio-cultural background, school, students and teachers. The aim of this study was to examine factors promoting the negative attitudes of female students toward the study of Mathematics in five selected Junior High Schools within the Fanteakwa District of Ghana. The researchers, therefore, used the sequential explanatory mixed method in order to generate the data from 125 respondents comprising of 120 female students and 5 mathematics teachers using a questionnaire, a semi-structured interview, and a focused group discussion. The result showed that female learners in the district exhibited negative attitudes toward mathematics as a subject; they developed fear and hatred for the subject, lacked self-confidence. The causative factors of these attitudes included sociocultural backgrounds; the lack of female role models, early marriage, and gender stereotypes. Home factors were the lack of parental support, illiterate parents, overburden household chores and poverty. Teacher factors included not creating the enabling environment for teaching and learning, the lack of teaching learning resources during lesson. The lack of reward pages for teachers and students, in-conducive environments were factors relating to school environment. The study concluded that students in the district had negative attitudes toward learning of mathematics; they considered the subject to be difficult and developed hatred for it. These were attributed to factors relating to home, socio-cultural, school, students and mathematics teachers. The study implied that the government of Ghana and policy makers should develop gender-friendly curriculum, sensitise parents to support their children’s education, train teachers to handle the subject well. The researchers, therefore, suggest that further studies on the topic should be undertaking using a larger sample or the study can be repeated in other districts and or the entire country so that a more holistic picture can be generated.