The soybean aphid ( Aphis glycines Matsumura) has become a major pest of soybean in North America since 2000. Seven aphid resistance sources, PI 71506, Dowling, Jackson, PI 567541B, PI 567598B, PI 567543C, and PI 567597C, have been identified. Knowledge of genetic relationships among these sources and their ancestral parents will help breeders develop new cultivars with different resistance genes. The objective of this research was to examine the genetic relationships among these resistance sources. Sixty-one lines were tested with 86 simple sequence repeat (SSR) markers from 20 linkage groups. Non-hierarchical (VARCLUS) and hierarchical (Ward’s) clustering and multidimensional scaling (MDS) were used to determine relationships among the 61 lines. Two hundred and sixty-two alleles of the 86 SSR loci were detected with a mean polymorphism information content of 0.36. The 61 lines were grouped into 4 clusters by both clustering methods and the MDS results consistently corresponded to the assigned clusters. The 7 resistance sources were clustered into 3 different groups corresponding to their geographical origins and known pedigree information, indicating genetic differences among these sources. The largest variation was found among individuals within different clusters by analysis of molecular variance.