This study aimed to evaluate the root canal real length (RL) changes due to the mechanical instrumentation use with different flaring magnitudes. After access cavity, 60 mesial root canals of mandibular molars were randomly separated in three groups: Hyflex EDM (HF; #25/.12, #10/.05 e #25/~), Reciproc Blue (RB; R25), and MTwo (M2; #10/.04, #15/.05, #20/.06 e #25/.06). The RL was defined as the apical limit, and 2.5% sodium hypochlorite irrigating solution was chosen. After the access cavity (RL 1), cervical flaring (RL 2), and complete chemical-mechanical preparation (RL 3), the RL was evaluated. The RL was evaluated by a blind examiner with the aid of a microscope (16x) placing the endodontic file stop at the coronary reference. When comparing length measurements, the RL was shorter before instrumentation than that after instrumentation. A reduction of 0.65 mm (HF), 0.61 mm (RB), and 0.48 mm (M2) was observed. However, among groups, no statistical differences were found (p > 0.05). Under the conditions tested, it can be inferred that all mechanical systems provoked RL variations, which emphasizes the need for constant verification of the odontometry, mainly before root canal obturation.