Human papillomavirus (HPV) vaccination protects against six types of cancer—cervical, anal, oropharyngeal, penile, vulvar, and vaginal. In the United States (U.S.), HPV vaccination coverage in college students remains low, especially in the Mid-South region, despite the highest risk of HPV infections and disease burden. However, few studies have assessed HPV vaccination among college students here. This study examined factors associated with HPV vaccination among college students in the Mid-South and explored preferred strategies for promoting vaccination. A mixed-methods design comprising a cross-sectional, self-report online survey and dyadic virtual interviews was conducted. Simple random sampling was performed to recruit a total of 417 undergraduate students aged 18–26 from March to May 2021; convenience sampling was performed to recruit three sex-matched dyads of a total of six (four female and two male) undergraduates from survey respondents who had not completed the HPV vaccine series in May 2021. Binary logistic regression analyses showed HPV vaccine knowledge and perceived barriers to vaccination were factors contributing to coverage for both female and male students, while perceived risks of HPV and vaccine hesitancy were factors only among female students. Findings from the qualitative content analysis identified college students’ perceived barriers to the vaccination at multiple levels and preferred strategies for vaccination promotion, corroborating the findings from the survey study. The findings provide implications that benefit the development of tailored interventions aimed at facilitating catch-up vaccination among college students in the Mid-South region. There is an urgent need for further research and the implementation of effective strategies that address the identified barriers and improve HPV vaccine uptake in this population.