In this paper, 41 unique wheat cultivars of Shandong Province were collected to construct DNA fingerprint database by SSR technique. The results showed that 69 alleles were detected with considerable polymorphism by 46 pairs of primers employed in the experimental design. For UPGMA cluster analysis using NTSYS, it was found that 41 kinds of materials could be divided into three groups with the threshold at genetic similarity of 0.68. This study indicates that (SSR technique) could effectively detect the genetic relationships among the various wheat cultivars, and it could meet the requirements for the construction of DNA fingerprint database for wheat, providing theory basis for collecting, storing, classifying, evaluating, and establishment of the core germplasm of wheat cultivars genetic resources of Shandong Province.