The five distinct species of Galium L. (Rubiaceae) were identified in this work utilizing the PCR methodology. In Iraq, species genotyping with the RAPD-PCR approach will be mainly focused on. Five primers (OPB18, OPC2, OPB11, OPC14, and OPC8) in Galium species yielded a total of 50 polymorphic amplified products from 120 bands, and the Jaccard's coefficient varied from 0.1081 to 0.3714. The dendrogram divided the investigated species according to the bivariate (1-0) data and genetic similarities utilizing the UPGMA cluster algorithm. Thus, it is possible to conclude that the RAPD methodology is useful for analyzing molecular Characterization and determining the links between different plant populations. Molecular genetic marker-based biodiversity analyses are valuable tools for reintroducing and conserving rare and endangered species. These species were classified utilizing phenotypic characteristics, and their accurate taxonomic names have been identified by building a phylogeny tree utilizing RAPD-PCR to assess for kinship. Members of Galium have tight relationships, as seen by the UPGMA-based dendrogram, which is a polymorphic association that suggests that every species has a distinct evolutionary tendency and identity.