This work explores how pedagogical content knowledge (PCK) on evolution was modified by two biology teachers who participated in a professional development programme (PDP) that included a subsequent follow-up in the classroom. The PDP spanned a semester and included activities such as content updates, collaborative lesson planning, and the presentation of planned lessons. In the follow-up part, the lessons were videotaped and analysed, identifying strategies, activities, and conditions based on student learning about the theory of evolution. Data were collected in the first round with an interview before the training process, identifying these teachers’ initial content representation (CoRe) for evolution. Then, a group interview was conducted after the lessons, and, finally, an interview of stimulated recall with each teacher was conducted regarding the subject taught to allow teachers to reflect on their practice (final CoRe). This information was analysed by the teachers and the researchers, reflecting on the components of the PCK, possible changes, and the rationale behind their actions. The results show that teachers changed their beliefs and knowledge about the best methods and strategies to teach evolution, and about students’ learning obstacles and misconceptions on evolution. They realised how a review of their own practices promotes this transformation.