X-linked microsatellites represent an efficient complement of autosomal and Y-chromosomal markers in forensic and kinship analysis. DXS337 (n=208), DXS101 (n=208), HPRTB (n=206), DXS8377 (n=220), and DXS7423 (n=213) were genotyped in male and female samples from a Western Mexican Mestizo population using singleplex systems and polyacrylamide gel electrophoresis. Genotype distributions did not deviate from Hardy-Weinberg expectations, and pairwise allele combination analysis was consistent with independent segregation for every marker (p>0.05). Allele frequencies were not different by gender. Differences in allele distribution with respect to Caucasian population data (DXS101, HPRTB, DXS8377, DXS7423) seem attributable to the native Mexican component. For the set of five markers, the combined power of discrimination and the probability of exclusion in paternity tests in trios were greater than 0.999. The present data reveal that the panel of five X-short tandem repeats analyzed is highly informative in forensic identity and parentage studies in Western Mexico.