It is aimed to reveal the factors that shape pre-service teachers’ (PSTs) online teaching practices in the scope of the technological pedagogical content knowledge (TPACK) framework. Ten science and nine mathematics PSTs who experienced the practical part face-to-face and the theoretical part online of the Teaching Practice 2 course participated in this study based on multiple case study designs. The researchers collected data through online lesson videos and TPACK in Online Teaching Survey. Descriptive statistics were used for the survey analysis, while the document analysis technique was through TPACK in Online Teaching Checklist for online lesson videos. The results show that PSTs have some problems, especially in the technological and pedagogical knowledge components of TPACK. Both groups do not have adequate knowledge about the assessment's technological tools because of giving it the most minor importance. They generally use standard technologies such as presentations or office programs. Their use of the same technologies for identifying and teaching the subject indicates their limited conceptualizations of technological pedagogical knowledge (TPK) and technological content knowledge (TCK). PSTs need improvements in promoting teacher-student interaction through interactive learning and assessment tools. Some recommendations are offered for teacher education programs.