A total of 100 SSR sequences were isolated and cloned by means of SAM (Selectively Amplified Microsatellite)techniques and another one was obtained by searching the NCBI and EMBL databases. There were 89 SSR sequences used for design of special primers. As a result, the primers were designed at 82 loci from 71 fragments. Forty-one special primers were synthesized, pairing with 5'anchored degenerate SSR primer, to detect 39 SSR loci. Fifteen of them amplified the corresponding SSR sequences and Other 11 SSR primer pairs amplified non-expected fragments. In the end, 21 polymorphic primer pairs were selected from 26 primer pairs which amplified clear and robust DNA fragment by using the genome DNA of 37 litchi germplasm materials, and 22 locus-specific SSR markers were obtained.