Plants have been used for the treatment of various diseases for thousands of years. Plants and plant-products are good sources of medications and provide raw materials for modern pharmaceuticals used for various ailments. Ocimum sanctum and Ocimum gratissimumis known to be an important medicinal plants from the ancient period in India. Thisstudy was carried out to investigate the phytochemical screening, antibacterial efficacy and molecular comparison of the leaf extracts of Ocimum sanctum and Ocimum gratissimum in various solvents like hexanes, methanol, ethanol, petroleum ether and aqueous extracts. To evaluate the inhibition of the pathogens with the plant extracts, solvent extracts of the selected plant species were screened for antimicrobial activity using the agar well diffusion method. The antibacterial activity was analyzed using six different bacterial strains (Escherichia coli, Bacillus cereus, Staphylococcus aureus, Klebsiella pneumonia, Aspergillus niger and Pencillium notatum). The methanolic and ethanolic extracts of crude leaf powder were found to be most effective against the pathogenic bacterial strains selected for the study, but the hexanes and petroleum ether extracts show less activity as compared to other extracts. The qualitative phytochemical analysis was studied and compared. The results reveal that Ocimum gratissimum has more phytochemical components and antibacterial activity as compared to Ocimum sanctum. The presence of the secondary metabolites signifies the potential of Ocimum gratissimumas a source of therapeutic agents and also can be used as a source for developing antimicrobial agents. The estimation of DNA was done by using Agarose Gel Electrophoresis and it was revealed that the Ocimum sanctum showed higher molecular weight than the Ocimum gratissimum