In this study, we identified the luminescent layers containing a significant amount of alginite in the Upper Jurassic–Lower Cretaceous Bazhenov Formation named “the alginite-rich layers”. Lithological and geochemical methods were used to determine distinctive features of these layers and to evaluate their impact on the total petroleum generation potential of the Bazhenov Formation. We have shown that the composition of the alginite-rich layers differs significantly from the organic-rich siliceous Bazhenov rocks. Rock-Eval pyrolysis, bulk kinetics of thermal decomposition, elemental analysis, and the composition of pyrolysis products indicate type I kerogen to be the predominant component of the organic matter (OM). Isotope composition of carbon, nitrogen, and sulfur was used to provide insights into their origin and formation pathways. The luminescent alginite-rich layers proved to be good regional stratigraphic markers of the Bazhenov Formation due to widespread distribution over the central part of Western Siberia. They can also be applied for maturity evaluation of the deposits from immature to middle of the oil window, since the luminescence of the layers changes the color and intensity during maturation.