Background <p>The fact that medical students lie in this age (17–31&#xa0;years) range and that they play a crucial role in delivering healthcare services makes their knowledge, attitudes and behaviors about blood donation (BD) important.</p> Objectives <p>The aim of this study was to examine the knowledge, attitude, and behavioral characteristics of medical students regarding BD.</p> Materials and methods <p>This study was carried out with a total of 396 students at Gaziantep University Faculty of Medicine from April 1st–15th, 2022 The sociodemographic characteristics of the students and their knowledge, attitudes, and behaviors about BD were interrogated.</p> Results <p>Among the 396 participants, 32.5% reported having previously donated blood, and 74.2% expressed a willingness to donate in the future. BD rates did not significantly vary by gender, religious beliefs, or year of education. However, students who had received training on BD were significantly more likely to have donated (<i>p</i> &lt; 0.001). The mean knowledge score about BD was 16.37 ± 5.43 (out of 30), with no significant difference between males and females. Knowledge levels were notably higher among fifth-year students and those who had received prior training (<i>p</i> &lt; 0.001). Despite a generally positive attitude toward BD, factors such as fear of contagion, lack of time, and misconceptions about donation during the COVID-19 pandemic negatively influenced participation.</p> Conclusion <p>It was determined that medical students exhibited a considerable lack of knowledge about BD. Training was shown to have a positive impact on their knowledge levels regarding BD. Furthermore, students expressed concerns and demonstrated insufficient understanding of the risk of COVID-19 transmission during BD. Providing up-to-date training on this topic is crucial to addressing these misconceptions and enhancing their knowledge.</p>