In this study, we selected 83 cultivars, including Hefeng 25, from 13 breeding units of Heilongjiang Province. We amplified 43 of 100 simple sequence repeats (SSRs), which distributed on 20 linkage groups of the soybean genetic map. In total, 157 polymorphic alleles were detected in 43 pairs of SSR primers. For each primer , 2 to 7 alleles (mean 3.65 alleles) were detected in all cultivars. Cluster analysis revealed most of the varieties bred by the same breeding unit with high genetic similarity. Pairwise genetic similarity coefficients ranged from 0.216 to 0.937 (mean 0.638 4), which indicated that the genetic basis of soybean varieties from Heilongjiang province were narrow. The genetic composition can be broadened by introducing diverse and distant cultivars as parents in soybean breeding.