We addressed the species taxonomy of five-toed jerboas (Allactaga, subgenus Paralactaga) in the Middle East by applying molecular markers (cytochrome b and a partial 16S rRNA). The study consisted of 17 specimens from eight localities in the Middle East, representing both species: Allactaga euphratica and Allactaga williamsi. The phylogenetic reconstructions yielded three highly divergent lineages, which failed to conform to the recent taxonomy of Paralactaga. The first lineage (williamsi lineage) encompassed all the samples of A. williamsi from Turkey and Iran and also the specimens of A. euphratica from Lebanon. The haplotypes of A. euphratica were arranged into two lineages, which showed strong geographic associations. One lineage contained samples from Harran in Turkey and from Iran, while all the samples from Syria clustered in another lineage. The pairwise Kimura twoparameter values suggested similar divergences between the three lineages and were within the range reported for a sister species of rodents. Our results point to a cryptic species in A. euphratica and also provide evidence of the expanded range of A. williamsi further south to Lebanon.