ABSTRACT: With unique structure, thermal, and mechanical properties, graphene oxide (GO) fascinates the scientific community. It is the thinnest known material in the universe and the strongest ever measured and use in many biological applications. The GO sheets were designed using a Gaussian 09 package of programs with a density function theory and were introduced in ultrasonic dispersion and deposition on glass substrate using distillation and a vacuum furnace. The characterization of the prepared GO is done using XRD, SEM, E-graph and UV-Visible spectroscopy. Atomic force microscopy shows the layers to be generally free of defects. scanning electron microscopy showed the dispersed phase to consist of small graphitic flakes. X-ray photoelectron spectroscopy show that ultrasound is the essence of enhancing chemical reaction rate. The crystallinity of the produced material was characterized using x-ray diffraction (XRD). The results show the XRD patterns different for samples grown at variable temperature and time .The results of testing MDR bacterial pathogens were to show bacteriostatic and bactericidal of GO against two types gram negative E. coli and gram positive S.aureus bacteria, from the figuring of the quantum chemical parameters, GO has few energy gap with rise performance to interact with other embracing species.