Formative research assessing human papillomavirus (HPV) vaccine readiness in Uganda was conducted in 2007. The objective was to generate evidence for government decision-making and operational planning for HPV vaccine introduction. Qualitative research methods with children, parents, teachers, community leaders, health workers, technical experts and political leaders were used to capture understanding of socio-cultural, health system and policy environments. We found low levels of knowledge about cervical cancer and HPV. Vaccination and its benefits were well-understood; respondents were positive about HPV vaccination. Health systems were deemed adequate for HPV vaccine delivery. Schools were identified as a vaccination venue, given high attendance by girls aged 10-12 years. Communication and advocacy strategies to foster acceptance should provide information on cervical cancer, HPV vaccine safety, and side effects. Policymakers requested further detail on costs. Introduction of HPV vaccine could be integrated into existing reproductive health and immunization policies.