Background: Many patients who require blood transfusion do not have timely access to safe blood and blood products. WHO states that at least 1% of the nation’s population should donate blood voluntarily to meet the basic requirements of blood. In India, there is a need for about 8 million units of blood every year. The need is predicted to increase by 5% every year. This study aimed to assess the knowledge, attitude and practices regarding blood donation among the general populace of a valley district in Manipur. Methods: A cross-sectional study was conducted among the general populace of a valley district in Manipur. Data was collected using a structured questionnaire through face-to-face interview and was summarized using descriptive statistics like mean, standard deviation and percentages. Association between variables of interest was analysed using chi square test. Conclusion: About one-fourth of the participants in this study had adequate knowledge (25.3%), favourable attitude (27.9%) and had donated blood at least once (27.6%). Males were more likely to have adequate knowledge regarding blood donation and to have donated blood compared to females. There was a significant association between educational qualification and religion with the level of knowledge, attitude and practice of blood donation.