To evaluate the utility of Y-STR data for DNA testing in two ethnic populations of Xinjiang province, a sample of 338 subjects (121 Kazakhs and 217 Uighurs) was tested. In the Kazakh and Uighur populations, the haplotype diversity was 0.868 and 0.996, respectively, and the discrimination capacity was 0.5950 and 0.8940, respectively. High numbers of singleton haplotypes were observed among Xinjiang Uighurs, but fewer were found in Kazakhs. Our results were also compared with geographically and linguistically close populations.