Biogenic nanoparticles are evolving as an important branch of nanomedicine which is cost effective and ecofriendly. The current study deals with the synthesis of silver nanoparticles using Ocimum sanctum leaf extract as reducing and capping agent. With increase concentration of AgNO3 (1-5mM), there was considerable increase in yield of silver nanoparticles The prepared nanoparticles were characterized using UV-visible spectroscopy, Fourier transform infrared spectroscopy (FT-IR), Transmission electron microscopy (TEM), X-ray diffraction (X-RD) and Dynamic light scattering (DLS).UV-visible spectroscopy showed maximum absorbance at 420 nm due to surface plasmon resonance of silver nanoparticles. FT-IR spectral analysis indicates the presence of water soluble phenolic compounds as reducing and stabilizing agents in the synthesis of silver nanoparticles. TEM analysis showed the presence of nearly spherical particles of size range 3-15 nm. Additionally, X-RD analysis revealed that the synthesized silver nanoparticles were anisotropic face-centered, cubic crystalline, having a size of 17.082 ± 5.83nm. DLS further support the formation of thermally stable silver nanoparticles, possessing negative surface charge potential. Thus, Ocimum sanctum leaf extract promotes rapid bioreduction of optimum concentration of silver nitrate resulting in a significant yield of silver nanoparticles which can be exploited commercially for its antioxidant activity.