Thalassemia is a hereditary hemoglobinopathies that results from the absence of a globulin chain and it is a very common problem in Southeast Asia. Like many other countries’ community based premarital carrier screening (PMCS) of thalassemia is extremely rare in Bangladesh. This study aimed to investigate knowledge, attitude, practice, and awareness towards PMCS among University students in Bangladesh. A cross-sectional study was conducted from January to March 2020 using a self-administered questionnaire which was distributed to 911 Jahangirnagar University students aged 18 to 24. The questionnaire was composed of five parts: personal information, knowledge, attitudes, practice, and awareness. On the basis of the answers, knowledge, attitudes, practice, and awareness towards PMCS were evaluated. About (88.80%) of the total participating students thought that PMCS is necessary before marriage. In the case of thalassemia carrier parents, 81.00% of students replied that prenatal screening is important before pregnancy. 610 students (66.95%) of the total 911 students considered that thalassemia can be caused by marrying between blood relatives. Most of the students (79.80%) supported that thalassemia screening should be an obligatory procedure and 91.98% would like to raise social awareness about PMCS and thalassemia. The students of Jahangirnagar University have good knowledge about PMCS program and a positive attitude but it is difficult to implement in real life without continuing practice. The outcome of this study suggested the necessity of increasing student’s awareness of Bangladeshi PMCS program.