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This study compared the cyclic fatigue and the canal transportation promoted by Twisted File (TF) Adaptive and Navigator EVO systems when used with 2 different motions.Forty mesiobuccal roots of maxillary molars were scanned by using micro–computed tomography imaging before and after root canal preparation with the 2 instrument systems used with 2 motions (adaptive and continuous rotation). Samples were divided into 4 groups: TFA, TF Adaptive instruments under adaptive motion; TFC, TF Adaptive instruments under continuous motion; NA, Navigator instruments under adaptive motion; and NC, Navigator instruments under continuous motion. Root canals were prepared until 35.04 instruments. Apical transportation was analyzed by using micro–computed tomography at 3 levels: 3, 6, and 9 mm from the apex. The cyclic fatigue tests were performed by using a custom-made device. Ten instruments of each brand were activated by using a 6:1 reduction handpiece powered by a torque-controlled motor using the preset programs “custom mode” and “TF Adaptive” to activate 25.06 and 35.04 instruments. Kruskal-Wallis and Dunn tests were used to assess canal transportation, centering ability, and canal volume. The Student t test was used to evaluate cyclic fatigue (P = .05).At 3 and 9 mm, the canal transportation and centering ability were similar in all groups (P > .05). At 6 mm, TFC presented higher canal transportation toward furcal region than NA and NC (P < .05). After canal preparation, TFA promoted great dentinal excision, presenting higher canal volume than NA and NC (P < .05). Higher cyclic fatigue resistance was observed under continuous than adaptive motion regardless of system or tip/diameter of the instrument (P < .05).Both systems can be used under adaptive or continuous rotation. However, the life span of the instruments was higher when used under continuous rotation. Small canal transportation occurred when mesiobuccal root canals from maxillary molars were prepared until 35.04 instruments.TFC presented canal transportation toward furcal region at 6 mm from the apex.Higher canal volumes were observed in TFA.Centering ability was similar in all groups.Files presented higher cyclic fatigue resistance under continuous than adaptive motion.Both systems can be used under adaptive or continuous rotation.