The extraction of neem oil from neem seeds (Azadirachta indica) has garnered significant interest due to its wide range of applications in medicine, agriculture, and cosmetics. This study aims to explore the extraction methods of neem oil and evaluate its antimicrobial properties. Neem seeds were collected, dried, and subjected to solvent extraction using n-hexane to obtain the neem oil. The extracted oil was then analyzed for its antimicrobial activity against common pathogenic microorganisms, including Escherichia coli, Staphylococcus aureus, and Candida albicans, using the disc diffusion method. The results indicated that neem oil exhibited significant antimicrobial activity against all tested microorganisms, with varying levels of inhibition. The oil showed the strongest antimicrobial effect against Staphylococcus aureus, suggesting its potential as a natural antibacterial agent. The antimicrobial efficacy was dose-dependent, with higher concentrations of neem oil leading to stronger inhibition zones. This study underscores the potential of neem oil as an effective, eco-friendly alternative to synthetic antimicrobial agents. Further research into its active components and applications in pharmaceutical and agricultural industries is recommended.