The second part of this review discusses the application of molecular methods in endodontic microbiology research for a comprehensive characterization of the microbiota associated with different types of endodontic infections. Despite their recent introduction in endodontic research, molecular methods have already given a significant contribution to the understanding of endodontic infections and the future holds the perspective of a still better refinement of the knowledge about these infections. Molecular methods have revealed a higher complexity of the endodontic microbiota than previously reported by cultivation approaches. In addition to detecting some cultivable species in increased prevalence, molecular methods have also expanded the list of putative endodontic pathogens by inclusion of some fastidious bacterial species or even uncultivated bacteria that have never been previously found in endodontic infections.