Summary Background Computer-assisted teaching is becoming increasingly more important to acquire new knowledge and skills in medical curricula. The consequence of gender-characteristic personality traits on academic performance in e‑learning examinations are difficult to forecast. This study investigated gender-related differences in examination behavior among undergraduate medical students taking a web-based quiz. Methods A total of 1315 4th grade medical students at the Medical University of Vienna completing the compulsory online moodle-based ECG quiz 2017/2018 were enrolled into this observational study. Individual data of examination behavior and quiz results as well as results of the final annual exam were extracted. Students were grouped into 10 strata according to academic performance. Variables between both sexes were compared using a nonparametrical test. Examination variables were correlated to performance. Results Of the total study population 686 (52%) were female and 629 (48%) were male. The time until the first attempt and number of attempts performed was comparable between both sexes, however female students spent more time on the first attempt compared to their male colleagues (1592 sec [Q1–Q3: 999–2536] vs 1405 sec [Q1–Q3: 828–2395], p = 0.002), suggesting a higher self-discipline and risk-aversity. There was no difference regarding quiz scores or final ECG examination scores between female and male students (p = 0.869 and p = 0.396). Students who accessed the quiz earlier and less time spent for the first attempt tended to perform better at the final examination (rs = 0.20, p < 0.001 and rs = −0.15, p < 0.001). Conclusions Gender-related differences in examination behavior already described for nononline based examinations are similarly observable in e‑learning. For this test, gender-immanent traits seem not to twist final examination results and impact academic performance.