Microbial mats, due to stratification of the redox zones, have the potential to include a complete N cycle; however, an attempt to evaluate a complete N cycle in these ecosystems has not been yet made. In this study, the occurrence and rates of major N cycle processes were evaluated in intact microbial mats from Elkhorn Slough, Monterey Bay, CA, USA, and Baja California Sur, Mexico, under oxic and anoxic conditions using 15N-labeling techniques. All the major N transformation pathways, with the exception of anammox, were detected in both microbial mats. Nitrification rates were found to be low at both sites for both seasons investigated. The highest rates of ammonium assimilation were measured in Elkhorn Slough mats in April and corresponded to high in situ ammonium concentrations in the overlying water. Baja mats featured higher ammonification than ammonium assimilation rates, and this, along with their higher affinity for nitrate compared to ammonium and low dissimilatory nitrate reduction to ammonium rates, characterized their differences from Elkhorn Slough mats. Nitrogen fixation rates in Elkhorn Slough microbial mats were found to be low, implying that other processes, such as recycling and assimilation from water, are the main sources of N for these mats at the times sampled. Denitrification in all the mats was incomplete, with nitrous oxide as the end product and not dinitrogen. Our findings highlight N cycling features not previously quantified in microbial mats and indicate a need for further investigations of these microbial ecosystems. IMPORTANCE Nitrogen is essential for life. The nitrogen cycle on Earth is mediated by microbial activity and has had a profound impact on both the atmosphere and the biosphere throughout geologic time. Microbial mats, present in many modern environments, have been regarded as living records of the organisms, genes, and phylogenies of microbes, as they are one of the most ancient ecosystems on Earth. While rates of major nitrogen metabolic pathways have been evaluated in a number of ecosystems, they remain elusive in microbial mats. In particular, it is unclear what factors affect nitrogen cycling in these ecosystems and how morphological differences between mats impact nitrogen transformations. In this study, we investigate nitrogen cycling in two microbial mats having morphological differences. Our findings provide insight for further understanding of biogeochemistry and microbial ecology of microbial mats.