The objective of this study was to analyze the influence of different methods of cervical prepare in determining the first file of the apical preparation of root canal and working length. Thirty mesiobuccal fully formed roots of upper first permanent molars humans were used and the root canals were initially cleaned with K-file #10. Then, the teeth were randomly divided into three groups according to the type of enlargement: group I: ProTaper S1, SX; group II: K3 Orifice Opener 25.10 and 25.08; and group III: technique with hybrid K-type hand files and gates glidden drills # 3 and # 2. The specimens were taken to a stereomicroscope to evaluate the adaptation of the first file in the dentinal walls at length established. The data were submitted to Kruskal-Wallis and Wilcoxon tests. The reliability of the apex locator methods was performed by Spearman test. All groups showed statistically significant difference in the length of root canals after preparation of the cervical third (p < 0.05). The level of agreement between the measurement and visual electronics was higher (r = 98.07%) before the cervical enlargement, compared to that reported after cervical preparation (r = 96.42%) (p < 0.0001). Based on the methodology, was possible to conclude that cervical and middle thirds prepare were important to determine the first file diameter and it did lead to a decrease in the working length. The electronic apex was effective to determine the working length.