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The aim of this study was to compare centering ability and transportation values of the Primary WaveOne Gold (PWOG; Dentsply Sirona, Ballaigues, Switzerland) instrument with or without different glide path techniques. Centering ability and transportation were also compared between the 3 different glide path preparation groups.The mesiobuccal canals of 60 extracted human mandibular molars (curvature angles between 25° and 35° and radii ≤10 mm) were selected and randomly divided into 4 groups with 15 canals each. Canals were negotiated to patency with a #8 K-file. Canal preparations were performed by a single operator using precurved #10, 15, and 20 stainless steel manual K-files (group KF); a #10 stainless steel manual K-file followed by PathFiles #1–3 (Dentsply Sirona) (group PF); a #10 stainless steel manual K-file followed by the WaveOne Gold Glider (Dentsply Sirona) (group WOGG); and no further glide path preparation (group NG). Micro–computed tomographic imaging was used to scan teeth before and after glide path preparation. The final canal preparation was performed with PWOG; after which, all 60 specimens were scanned again using micro–computed tomographic imaging. Canal transportation and centering ratio values were determined over levels 3 mm, 5 mm, and 7 mm from the root apex after glide path preparation and again after final preparation with PWOG. One-way analysis of variance was used to statistically compare groups. The significance level was set at P < .05.No statistically significant difference in the mean centering ratios was found after glide path preparation when the KF, PF, and WOGG groups were compared (P > .05). The mean combined transportation values after glide path preparation were statistically significantly higher in the KF group compared with the PF and WOGG groups (P > .05). There was no statistically significant difference in the mean combined centering ratios or transportation values of the NG and 3 glide path groups in combination with PWOG over the 3 levels (P > .05).KF group showed the highest mean transportation values during glide path preparation. However, the PWOG instrument's performance regarding centering ability and transportation was not influenced by the different glide path/no glide path preparation groups.We compared canal shaping with the Primary WaveOne Gold instrument after either no glide path preparation or glide path preparation using 3 different techniques.There was no statistically significant difference in the mean centering ratios after glide path preparation with the WaveOne Gold Glider, K-files, or PathFiles.The mean combined transportation values after glide path preparation were statistically significantly higher for K-files than for PathFiles and the WaveOne Gold Glider.There was no statistically significant difference in the mean combined centering ratios or transportation values of the no glide path group and the 3 glide path groups in combination with the Primary WaveOne Gold instrument at the 3-mm, 5-mm, and 7-mm levels.