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The TPACK framework is one of the most prominent frameworks for describing teacher knowledge for successful use of digital technologies in teaching. This study addresses several theoretical and methodological issues, which have often been criticized in TPACK research, like lack of conceptual clarity, lack of subject-specificity and lack of evidence of reliability and validity. Therefore, a biology-specific knowledge self-report and performance assessment were developed and evaluated. Both test formats addressed the four technology-related dimensions of TPACK (T-dimensions: TK, TCK, TPK, TPACK). 206 biology teacher students from an Austrian university participated. The study aimed at examining the levels of discrimination and convergence among the T-dimensions and analyzing the internal structure of the T-dimensions assessed by the two assessments.