Bioinformatics is a subject that analyzes and interprets the collection, processing, storage and dissemination of biological information, and explores biological problems by using information technology and related means. The outbreak of novel coronavirus pneumonia has been nearly three years. With the advancement of vaccine research and development in various countries, mass vaccination is bound to usher in a new turning point for epidemic control. China’s policy of free vaccination for the new coronavirus vaccine will undoubtedly improve the vaccination rate, however, the vaccination rate still depends on people’s will. This paper takes college students as the main research object. In the context of bioinformatics analysis, a questionnaire survey was conducted based on their knowledge, attitudes, and practice levels of COVID-19 vaccines. The results vividly illustrate five conclusions. Firstly, the reliability of information sources positively affected college students’ cognitive level, degree of trust and inoculation intention of COVID-19 vaccination. Secondly, the professional degree of media positively affected college students’ degree of trust and inoculation intention behavior of COVID-19 vaccine. Thirdly, the cognitive level and degree of trust about COVID-19 vaccine positively affected college students’ intention behavior of COVID-19 vaccine vaccination. Fourthly, the positivity of information positively affected the willingness of college students to vaccinate against COVID-19 vaccine. Fifthly, there were significant differences in the willingness and behavior of college students with different genders in COVID-19 vaccination. Finally, three suggestions are given to improve college students’ willingness to vaccinate against the coronavirus and to cope with similar incidents in the future.