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Background: The root canal anatomy is a complex space, and using a single-cone obturation technique does not guarantee a complete filling. Aim: This research aimed to evaluate the actual temperature of the biosealer during the obturation phase inside the root canal using a new hot technique. In addition, this study evaluated the penetration depth of the biosealer inside dentinal tubules. Setting and Design: The current study was concluded on two ex vivo experiments; the former tested the temperature evaluation. The latter examined the penetration depth of the biosealer. Materials and Methods: In this study, 42 sec mandibular premolars were used, and two experiments were carried out. In the first experiment, two premolars were utilized, and two thermocouples of K-type for each tooth were used. During the second experiment, the penetration depth of the biosealer was examined, 40 teeth were selected. Root canals were prepared then divided into two obturation groups: A, gutta-percha, and biosealer with single-cone technique and B, gutta-percha, and biosealer with a newly proposed obturation method. Confocal laser scanning was used to assess the penetration of the sealer. Statistical Analysis Used: In the first experiment, the Bonferroni method was performed to compare the temperature data. The tests used in the second experiment were Shapiro–Wilk's test, Kruskal–Wallis, and Mann–Whitney tests. Results: The first experiment results showed that the heat does not reach the apical third using the new obturation method. In the second experiment, the results showed in Group B significantly higher biosealer penetration into the dentinal tubules as compared to Group A (P < 0.05). Conclusions: The highest level of penetration of the biosealer in the dentinal tubules was observed in the group of the new obturation method, and the last apical 3 mm remained at 37° using this novel hot modified technique providing no risk of chemical alteration of the biosealer. [ABSTRACT FROM AUTHOR]