This study aims to explore public perception of blood donation and assessment of its impact on health service delivery in Juba County of South Sudan. A positivist and interpretivists philosophy were adopted, utilizing quantitative and qualitative data collection through questionnaires. A pilot study ensured questionnaire reliability. The study’s population includes all blood donors in Juba County. Questionnaires and interviews were the main instruments for the data collection. The purposive and convenient sampling technique were adopted whilst the Krejcie and Morgan's (1970) ^[27] formula was used in calculating the sample size of 267 to represent the population. Data analysis was carried out by the use of Statistical Package for Social Science (SPSS) version 21 and thematic analysis. Findings of the study revealed mixed level of awareness and knowledge about blood donation among the public in South Sudan with low level of awareness and knowledge recorded a mean score range of 1.00-2.24, average level of awareness and knowledge recorded a mean score range of 2.25-3.49, a high level of awareness and knowledge recorded a mean score range of 3.50-4.74 and a very high level of awareness and knowledge recorded a mean score range of 4.75-6.00