Abstract: Simple sequence repeat (SSR) markers have been used extensively in studies on the heredity of flora and fauna because they are widely distributed throughout the genome, codominant, stable, and polymorphic. The genomic DNA of Potentilla anserina was digested with the restriction enzymes EcoRⅠ and MseⅠ and the digested fragments were hybridized with biotinylated (AG)₁₅ and (GT)₁₅ probes. The fragments containing SSR genomic loci were adsorbed onto magnetic beads coated with streptavidin. Then, by elution, PCR amplification, cloning, and sequencing, a collection of SSRs was obtained. A total of 236 effective clones was acquired. Eighty sequences were randomly selected for sequencing, and 68 of them (85%) were found to be single sequences. Forty sequences including SSR loci were selected and 40 primers were developed using primer design soft (Primer 5). Finally, 20 pairs of primers with rich polymorphism and stable amplification were obtained through preliminary screening. The success rate of amplification was 50%. A preliminary analysis using the 20 pairs of primers amplified 338 polymorphic sites, with an average of 17 sites per primer pair. The mean values of effective allele number (Ne), polymorphism information content (PIC), Nei’s gene diversity (H), and Shannon-Weaver index (I) were 1.2440, 0.8996, 0.1768, and 0.3078, respectively. These 20 pairs of SSR primers provide novel molecular markers for research on the genetic diversity of P. anserina.