A simple method for isolation of microsatellites from the Japanese squirrel,Sciurus lis,without constructing a genomic library

We developed a simple and easy method to isolate microsatellites without screening genomic libraries by hybridization. The method requires only three basic techniques: polymerase chain reaction, DNA cloning and sequencing. We applied this method to develop microsatellite markers for the Japanese squirrel and isolated 45 clones that contained repetitive sequences. Among the 22 clones that we tested further, we found 11 diagnostic microsatellite loci that are applicable to the molecular ecological study of Japanese squirrels.