We describe the isolation, development and application of six microsatellite loci in the song sparrow, Melospiza melodia, their amplification in related species and problems encountered in genotyping from high quality DNA. Heterozygosity in an island song sparrow population was high (> 70%) for five out of six loci. Small scale cross-species amplification showed up to three alleles per locus for other Emberizidae. Relatively high levels of allelic dropout were observed with two loci when using standard template DNA. These problems could be partially alleviated by diluting template DNA, and could always be eliminated by repeated genotyping.