The most dominant cause of root canal treatment failure is nonhermetic obturation. It could be prevented with the use of sealer material. The sealer that has a good bond with the dentin wall is sealer epoxy bis-phenol resin sealer and Glass Ionomer Cement. A natural material that could dissolve guttapercha in root canal re-treatment is eucalyptus oil. The eucalyptus oil has an antimicrobial effect, bio-compatible and recommended for use in root canal retreatment. Objective: To determine the effect of epoxy bis-phenol resin sealer and Glass Ionomer Cement sealer in softening the gutta-percha using cajuput oil. Methods: The design of this study was pure laboratory experimental. This study was using 24 samples of single root canal acrylic block. The samples were treated differently by the type of sealer used and incubation time (15 days, 22 days, 29 days). The data were processed using one-way ANOVA and Post-Hoc Tukey test with significance level 95% (p <0,05). Result: There was a significant effect of epoxy bis-phenol resin sealer and Glass Ionomer Cement sealer in softening gutta-percha using cajuput oil. The result of one-way ANOVA analysis obtained a significance value of 0.000 (p <0,05). The results of Post-Hoc test analysis showed that the resin sealer which was incubated for 15 days after contact with cajuput oil for 5 minutes had a higher softening value compared to the Glass Ionomer Cement sealer which was incubated for 15 days, 22 days and 28 days after contacting with cajuput oil for 5 minutes. Conclusion: Sealer epoxy bisphenol resin is easier to remove than Glass Ionomer Cement sealer with cajuput oil solvent.