Multilocus approaches are essential for accurately recovering the evolutionary processes underlying species and population history. However, historical inferences in non-model organisms are still almost exclusively based on mitochondrial DNA due to the difficulty of obtaining multiple informative loci. Here, we use a genomic library based approach, to generate 15 novel anonymous nuclear markers (ANMs) from the ground spider Harpactocrates globifer Ferrández 1986. The ANMs cross-amplify and sequence well in the target species and its two closest relatives, and some of them also in the more distantly related H. ravastellus Simon 1914. Levels of nucleotide diversity of the ANMs within H. globifer ranged from 0.05% to 1.4% and average sequence divergence between close congeneric relatives from 0.02% to 13.9%, supporting the utility of these loci in population and species-level analyses. Moreover, a cross-species amplification screened in other spider taxa showed that some of the loci could also potentially be useful in more distantly related genera.