"Background: Blood and blood components save lives in various conditions. In India there is a need of about 8 million units of blood per year and only one-third of this is obtained from voluntary donors. Assessment of awareness and attitude regarding blood donation will help in designing effective health education strategy to improve blood donation. Objectives: To assess the knowledge and attitude regarding blood donation among individuals aged 18-60 years in rural Puducherry. Methods: A cross-sectional survey using a structured and pre-tested questionnaire in the rural service area of Puducherry. Results: Out of 288 respondents, 229 subjects (79.5%) were aware that blood could be donated, and only 14.8% of them knew about the correct frequency of blood donation. Around 80% of these subjects felt that the victims of road traffic accident required blood transfusion. Among those who were aware that blood could be donated, 40 subjects (17.5%) had donated blood in past. Most of them had donated blood for their relatives (55%). Among non-donors the most common reason for not donating blood was “never considered” (34.2%). However, three fourth of the non-donors have shown their willingness to donate blood in future. Electronic and print media were found to be the most common source of knowledge. Conclusion: Though the awareness regarding blood donation was high, the practice of voluntary blood donation was remarkably low. Education and motivation through various media is recommended to eliminate misbelieves and to reinforce positive attitudes towards blood donation."