The content knowledge tutors possess during chemistry sessions in the implementation of gender-responsive pedagogy was identified in this study. Eighteen (18) research participants made up the study's population. The study employed non-probability sampling approaches. For the study, quota sampling and a purposive sampling approach were used. Two (2) chemistry tutors and a vice-principal of the college were chosen using a purposeful sampling approach. Twenty percent (20%) of 38 females and twenty percent (20%) of 34 males were chosen for the study using quota sampling. The major data-gathering instruments were a self-constructed interview guide and structured observation checklists. Thematic content analysis was used to examine the qualitative data. The paper finds that St. Ambrose College of Education instructors displayed knowledge and awareness of gender pedagogy during chemistry sessions based on the findings. In many ways, this conclusion has policy implications. Gender-responsive pedagogy, for example, should be encouraged across the curriculum in all subject areas to expand its practice among teachers at the College. Mentors and Tutors should be mandated to apply gender-responsive pedagogy in their teaching in the continuous capacity-building programme organised by the Ghana Education Service and the Ministry of Education, by consciously including its application in the basic school and Colleges of Education curriculum.
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