I collected mitochondrial DNA sequences—hypervariable region 1 of the control region—from 281 eastern chimpanzees in 19 geographically defined populations and calculated genetic diversity measures to test the hypothesis that populations inhabiting the reconstructed locations of Pleistocene forest refugia harbor higher genetic diversities than those of other populations. The hypothesis is only weakly supported. Population genetic diversity is not significantly correlated with geographic proximity to refugia, with the area of forest that the populations currently occupy, or with the degree of geographic isolation of the populations. However, the two populations displaying the consistently highest genetic diversities are located in refuge areas: Uganda's Rwenzori Mountains and the eastern Democratic Republic of Congo's Ituri Forest. These results, in combination with previously findings, imply that chimpanzees may have lived both in and out of refugia during periods when tropical forests were restricted to refugia. This interpretation is consistent with the notion of chimpanzees as an extraordinarily vagile species, capable of maintaining gene flow across habitat mosaics of forest, woodland, and savannah.