• The characterization of Capsicum spp. present in germplasm banks helps in the development of new cultivars. • ISSR and SSR molecular markers are tools that show efficiency and effectiveness in the detection of genetic variation, their use allows the reduction of maintenance costs of Germplasm Banks through the elimination of duplicate material/genetic duplicates. • SSR markers showed greater hiding power and consequently greater detection of polymorphisms in Capsicum spp. Molecular markers are important tools to study genetic diversity, guiding characterization practices within the genus Capsicum. The objective was to characterize 26 Capsicum spp. genotypes using 13 molecular markers ISSR and 17 SSR. The genetic material for the study came from the collections of Capsicum spp. preserved at the Universidade Federal do Maranhão and the Instituto Federal do Espírito Santo, Campus in Alegre. The polymorphism of primers was analyzed from two dendrograms generated by the hierarchical method UPGMA, and their entanglement was obtained using the Tanglegram algorithm of the den extend package of the R software. The 26 Capsicum genotypes were structured in four and six groups of genetic diversity using the ISSR and SSR primers , respectively. Both markers were efficient to detect diversity among genotypes. No duplicates were found among the genotypes evaluated, showing genetic materials important both for diversity conservation and the development of new Capsicum spp. cultivars.