This study was to determine the frequencies and parameters of 26 Y chromosomal short tandem repeat loci of Han population from Zhoukou in Henan province. Private alleles of each area were also studied to better explore the application value of DNATyperY26 kit. Blood samples of 802 nonrelated male were collected and genotyped through PCR amplification and capillary electrophoresis based on DNATyperY26 kit. Genotype data was then analyzed for allele frequencies and parameters. We found 180 alleles with their frequencies ranging from 0.001~0.854. Genetic diversity (GD) value of Y-STR loci distributed from 0.261~0.965 with most of them higher than 0.5. Discrimination capacity (DC, 0.969) of the kit was very high since 777 haplotypes were detected in 802 samples. The study also showed high haplotype diversity (HD, 0.9999). Moreover, we observed 5, 5, 10, 7 private alleles respectively for each area in Zhoukou. In conclusion, DNATyperY26 kit can be widely applied to the research of forensic science and population genetics. Analysis of private alleles may benefit in source region inference, which still needs further exploration.