Plants have served human kind as sources of medicinal agents since its earliest beginnings. In fact natural product once served as the source of all drugs. OcimumSanctumL. is an aromatic plant in the family Lamiaceae. The main chemical constituents of Tulsi are: Oleanolic acid, Ursolic acid, Rosmarinic acid, Eugenol, Carvacrol, Linalool, and β-caryophyllene, have been used extensively for many years in food products, perfumery, and dental and oral products. Recent studies suggest that Tulsi may be a COX-2 inhibitor, like many modern painkillers, due to its high concentration of eugenol. The present study was to evaluate the qualitative estimation of phytochemicals and antimicrobial activity ofaqueousand methanol extracts of root and leaves of Ocimum sanctumagainst pathogenic bacteria i.e. Escherichia coli, Proteus mirabilis, Staphylococcus aures.Study has been shownthe presence of steroids, alkaloids and tannins. Significant antimicrobial activity of plant extract has been observed.