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Botswana was amongst the first African countries to introduce universal access to HIV testing through voluntary counseling and testing (VCT) and routine HIV testing (RHT). However, despite years of the implementation of VCT and RHT, in 2013, only 70% of Batswana had ever been tested for HIV because of the limitations of the testing options. Alternative HIV testing options such as HIV self-testing (HIVST) have not been adopted, and its feasibility and acceptability have not been determined. We assessed the acceptability of and opinions of tertiary students at the Institute of Health Sciences in Lobatse about HIVST. Five focus group and five in-depth interviews were conducted with 45 students. Thematic data analysis using NVivo 10 were conducted. The findings revealed that the HIVST was acceptable to the students who believed that adopting it as an HIV testing approach would address an unmet need for testing. They would utilize HIVST because it would assure confidentiality and privacy, it is convenient and offers the flexibility of when and where to conduct the self-test. However, there are were also concerns about the cost of self-testing kits, the lack of post-test counseling, and the lack of strategies to link the self-test results to HIV programming. They recommended that the concerns should be addressed to make HIVST effective and that potential users of HIVST should be provided with clear instructions on how to self-test. HIVST is an acceptable testing option that could complement available HIV testing and counseling services provided in public health facilities.