The present study aims to determine pre-service biology teachers' self-confidence levels concerning their technological pedagogical content knowledge with respect to variables namely class level and computer knowledge. Designed as a survey, 91 pre-service teachers enrolled in the biology education programme participated in the study. Data were collected by using the Technological Pedagogical Content Knowledge Self-Confidence Scale developed by Graham, Burgoyne, Cantrell, Smith, Clair and Harris (2009) and adapted in Turkish by Timur and Tasar (2011). Findings of the study show that pre-service teachers have a high self-confidence concerning their technological pedagogical content knowledge. It was also found that there was no significant difference in pre-service teachers' self-confidence with respect to the class level. However, it was found that pre-service teachers who had enrolled in a computer course during their education had higher self-confidence.