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National-level data suggest that sexually transmitted infection (STI) testing rates among young adults are low. The purpose of this study was to evaluate the acceptability of an STI self-testing program at a university health center. Few evaluations on the acceptability of collegiate self-testing programs and their effect on testing uptake have been conducted.To assess acceptability and uptake of self-testing (urine and self-collected vaginal swab), we conducted a brief self-administered survey of students accessing a large US-based university health center from January to December 2015.In 2015, University Health Services experienced a 28.5% increase in chlamydia (CT)/gonorrhea (GC) testing for male individuals and 13.7% increase in testing for female students compared to 2013 (baseline). In 2015, 12.4% of male students and 4.8% of female students tested positive for CT/GC via clinician testing, whereas 12.9% of male students and 12.4% of female students tested positive via self-testing. Female students were more likely to test positive for CT/GC when electing to test via self-test versus a clinician test (χ2(1, N = 3068) = 36.54, P < 0.01); no significant difference in testing type was observed for male students. Overall, 22.5% of students who opted for the self-test option completed the acceptability survey; 63% reported that their main reason for testing was unprotected sex. In the past year, 42% reported 4 or more partners. The majority were very satisfied and likely to use the service again (82%).Self-testing may be an efficient and effective way to provide STI testing for students and increase testing uptake. Self-reports of multiple partners, unprotected sex, and detected infections suggest that at-risk students are using the service.