Background: The purpose of this study was to compare the quality of the coronal seal of three root canal sealers and two obturation techniques using the bacterial penetration method. Methods and Results: A total of 132 single-rooted human teeth with fully developed apices were used. The teeth were randomly assigned to three experimental groups according to the endodontic sealer used. Group 1: Samples (n=44) were obturated using a zinc oxide eugenol-based sealer, Pulp Canal Sealer EWT. Group 2: Samples (n=44) were obturated using an epoxy resin-based sealer, AH Plus. Group 3: Samples (n=44) were obturated using a bioceramic-based root canal sealer, Well-Root ST. Each group was subdivided into 2 equal subgroups in accordance with the obturation technique being used: CLCT and ThOT. Thus, 6 subgroups were formed: Sub-1A: Pulp Canal Sealer/CLCT; Sub-2A: AH Plus/CLCT; Sub-3A: Well-ROOT ST/CLCT; Sub-1B: Pulp Canal Sealer/ThOT; Sub-2B: Ah Plus/ThOT; Sub-3B: Well-ROOT ST/ThOT. A dual-chamber device was used to evaluate bacterial leakage. Fresh medium and E. faecalis were added to the upper chamber every 4 days. The broth was monitored for color change daily for 33 days. Significant differences were found among Sub-2A vs. Sub-1B (P=0.023), Sub-1A vs. Sub-3A (P=0.014), Sub-1A vs. Sub-2B (P=0.024), Sub-1A vs. Sub-3B (P=0.002), Sub-3A vs. Sub-1B (P=0.003), Sub-2B vs. Sub-1B (P=0.005), and Sub-1B vs. Sub-3B (P<0.0001). There was no significant difference in the average occurrence of turbidity between CLCT and ThOT (P=0.718) Conclusion: Regardless of the obturation technique, all root canal sealers exhibited leakage; however, the bioceramic-based root canal sealer appeared to perform better than the epoxy resin-based sealer and the zinc oxide eugenol-based sealer.