The AmpFlSTR Y-filer kit, including 16 Y-STRs was analyzed in 326 males from Chihuahua, at North Central, Mexico. Allele frequencies and gene diversity for each locus were estimated. Four allele duplications, namely DYS389II, DYS390, DYS391 and DYS439; and one allele null at DYS448 were observed in the sample. The haplotype diversity was 99.97+/-0.3%. The AMOVA results, including a previous report from West of Mexico (Jalisco), showed that most of the genetic variability between these Mexican populations is attributable to intrapopulational differences (99.87%). This result supports a low-genetic differentiation between males from North and West regions of Mexico.