Pieces of evidence have continued to emerge, demonstrating the extensive efficiency and effectiveness of the DNA database in assisting criminal investigations around the world. Therefore, the present study aimed to determine the awareness level on the prominent role of Forensic DNA Database on Crime Investigation in Nigeria: a case study of Benin City. In conducting this research, a total of 458 questionnaires were distributed around Benin City between the periods of 12th January 2020 to 21st March 2020, with a particular focus on security agents and students. The questionnaire comprised of three main categories: Socio-demographic characteristics, Information about the National Forensic DNA Database, and Information about DNA evidence, and Nigeria Criminal Justice system. For the analysis of data collected; the statistical tool used was also Statistical Package for Social Sciences, version 22 for windows. Responses were compared using chi-square and presented as counts and percentages. In determining the level of awareness, the following responses were obtained. Of the total population: 53.28% had no idea about forensics, 19.21% were uncertain and 27.54% knew about forensics. The same trend was observed with Forensic DNA profiling, 42.14% did not know, 22.27% were uncertain and 35.59% demonstrated good knowledge of Forensic DNA profiling. On the knowledge about the National Forensic DNA Database, 48.47% had no knowledge, 22.27% were uncertain and 29.26% were knowledgeable about it. The result of the present study revealed that the awareness level of the forensic DNA Database was found to be inadequate.