Kuhl's pipistrelle is found from Europe and North Africa all of the way to Asia, yet studies have thus far concentrated on the western limit of its distribution. Here we form a multi-marker picture of the diversity of Kuhl's pipistrelle at a mid point in the Arabian peninsula in an attempt to redress the western sampling bias and to represent a region from which no genetic data has thus far been presented for this species. The three Arabian Cytochrome b haplotypes showed a clear divergence of 19 substitutions from those found in either Europe or North Africa. Molecular dating suggests the Arabian population split from the remaining Kuhl's somewhere between 0.7 and 1.7 million years before present around the time of a series of aridification events across northern Africa. Well supported lineages within Arabia are typical of that which may be seen after an expansion from multiple Pleistocene refugia, but may also reflect the loss of intermediate haplotypes during historical population fluctuations. A long-term population contraction coincides with climatic changes towards those conditions more typical of contemporary Arabia.