Molecular markers are the basic tools of genetic research， which are widely used in genetic diversity research， germplasm identification， genetic map construction and gene mapping. In this study， Barley SNP 50K chip was used to detect SNP in barley core resources with wide genetic background， and a series of dimorphic SNPs with high polymorphism were selected， and 124 KASP markers were developed. 43 barley germplasms from different geographical sources were used to test their effectiveness， and 56 KASP markers were initially screened out. Eighteen high-quality KASP markers were selected based on MAF （minor allele frequency）≥0.40 and PIC （polymorphic information content）≥0.45 criteria and applied to construct phylogenetic trees of 98 Jiangsu barley varieties. The results showed that barley materials with the same geographical origin and close relatives could be grouped into one class. These results indicate that KASP markers have guiding significance and application value in barley variety identification genetic relationship analysis of barley resources and population division. At the same time， the SNP fingerprint of 98 Jiangsu barley varieties was constructed， which verified the feasibility of KASP technique in barley varieties identification in China. The developed KASP marker can identify barley varieties accurately and quickly ， which is helpful to the scientific and standardized management of barley germplasm resources and the study of genetic diversity.