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The study was designed to test application of ultraviolet light to root canal walls, as a mean of complementary immediate disinfection after the use of sodium hypochlorite.Root canals were infected ex vivo with Enterococcus faecalis for 48 hours. Nonattached bacteria were washed away, and the remaining attached bacteria were subjected to disinfection, with 5% sodium hypochlorite alone or followed by exposure to ultraviolet light (254 nm, 300 mJ/cm2). Root canals were then tested for remaining viable bacteria. Canals were obturated and tested again after 14 days.Sodium hypochlorite alone achieved negative cultures in only 47% of the cases, but 96% was achieved with sodium hypochlorite followed by ultraviolet light (P < .001). This status was also maintained after 14 days.Illumination of root canals with ultraviolet light may be an effective supplementary means to achieve immediate disinfection of infected root canals.