A genomic library enriched with (gT)(n) repeats from tartary buckwheat (Fagopyrum tataricum) was constructed using 5'-anchored PCR for the development of microsattellite markers. Sequencing analysis of 5 clones from the library showed that they all contained microsatellites (totally 10 loci), and each was unique. An additional locus-specific primer was designed according to flanking sequence. Two of the microsatellite loci of 10 tartary buckwheat varieties were amplified using an anchored primer and a locus-specific primer, which revealed a clear polymorphic pattern. The data confirmed that the degenerate primer was reliably anchoring at the 5'-end of the microsatellite, and the primers developed based on this technology could be used for diversity analysis of tartary buckwheat.