An all-inclusive approach: A universal protocol for the successful amplification of four genetic loci of all Onscidea

Accounting more than 3,700 described species, Oniscidea is the largest and at the same time the only terrestrial isopod suborder inhabiting almost all terrestrial biomes. Despite the great effort dedicated on describing taxonomic diversity of Oniscidea, mainly employing morphology, there is still a considerable number of species/genera of uncertain generic/familiar assignment. Based on different morphological characters, alternative evolutionary relationships have been proposed to describe the diversity of Oniscidea at different phylogenetic levels. Accumulating morphological and genetic data are repeatedly challenging the monophyly of established taxa, undermining the validity of several morphological characters traditionally used in terrestrial isopod taxonomy, leading to often revisions of the current taxonomy of the Oniscidea . The use of genetic data facilitates the efforts to reconstruct the complex evolutionary history of the focal group by providing important data for the identification, delimitation, and description of species. The proposed protocol with universal PCR conditions and primers was used to successfully amplify COI, 16S, 28S and NAK loci in diverse Oniscidea taxa. The application of this protocol is anticipated to facilitate the generation of new genetic data and hence promote scientific research in Isopoda taxonomy, evolution, ecology, and other related fields.