This study was designed to evaluate the knowledge, attitudes, and perceptions of non-healthcare students regarding antibiotic use and antibiotic resistance at Central University, Miotso, Ghana. This study involved students from all levels studying non-healthcare courses at the Central University of Ghana. The study was conducted over three months. The questionnaire consisted of demographics as well as questions on respondents’ knowledge, attitudes, and perceptions regarding the use of antibiotics and antibiotic resistance. Data were analyzed using STATA analysis software version 12.0. The sample size comprised 1000 students, with 550 being female and 450 male. Of these, 60.96% stated that they had used antibiotics within the last 12 months. A small number of respondents (31%) indicated that they always consulted with a physician before starting an antibiotic, and students (68.75%) across all levels agreed that most colds get better on their own without antibiotics. This study also showed a significant association between students’ knowledge of antibiotics and their level of study (p = 0.007). Our study suggests that most of the respondents were aware of the severity of antibiotic resistance, but almost all of them had inadequate knowledge on the use of antibiotics and on how they can contribute towards curbing the menace of antibiotic resistance. There is therefore a need to include information programs on the judicious use of antibiotics and antibiotic resistance as part of all degree programs.