Introduction: The plant kingdom offers an infinite and therapeutic unexploited source of medicines. Various parts of some plants viz. fruits, flowers, bark, stem and roots, are seen to possess enormous bactericidal and fungicidal potential. Aim/objective: The aim of the study was to evaluate the in vitro-antimicrobial activity of Annona squamosa Linn. leaf extract. Methods: In-vitro antimicrobial activity of the sequential leaf extracts of Annona squamosa Linn. in different solvents like petroleum ether, chloroform, ethyl acetate, acetone, ethyl alcohol, methyl alcohol, and water, was studied against various gram-positive __ampersandsignamp; gram-negative bacterial strains using zone of inhibition. Agar well diffusion method __ampersandsignamp; Agar disc diffusion methods were selected to evaluate the antibacterial efficacy and the minimum inhibitory concentration (MIC) of all the extracts was done using Agar well diffusion method. The reference Antibacterial antibiotics viz., Chloramphenicol __ampersandsignamp; Ampicillin; and Antifungal antibiotics viz., Nystatin __ampersandsignamp; Clotrimazole were also tested against these standard microorganisms used in the assay and the results were compared with that of the plant extracts. Results: The study results revealed that all the seven sequential Annona squamosa Linn. leaf extracts showed significant bactericidal and fungicidal potential against the microorganisms under study. Highly polar solvents i.e. ethyl alcohol, methyl alcohol, and water showed the most significant antibacterial and antifungal activity against all tested organisms. Conclusion: It can thereby be concluded that the plant material under study can be considered as a possible source of various phytochemical constituents having an in vitro antimicrobial potential.