Introduction: For the complete removal of the intracanal debris and the infectious agents, the intracanal irrigation plays a significant role. Hence, in our study, we assessed the efficiency of diode laser in root canal disinfection. Materials and Methods: We conducted an in vitro study on 45 extracted single noncarious teeth. We equally divided the teeth into controls, Endovac, and diode laser. Later the mean colony-forming units (MCFU) of the Enterococcus faecalis were calculated and compared statistically. Results: We observed that the MCFU in the controls, Endovac, and diode laser was 10 8 /ml, 10 4 /ml, and 0, respectively, after the incubation. Conclusion: We can conclude that diode laser can be successful in irrigation of the root canals.