To perform whole genome scanning for complex trait analysis, we isolated and characterized a total of 21 novel genomic-SSRs and EST-SSRs for yellow perch (Perca flavescens), using the methods of construction of SSR-enrichment libraries and EST database mining of a related species P. fluviatilis. Of 16 genomic-SSR primer pairs examined, eight successfully amplified scorable products. The number of alleles at these informative loci varied from 3-14 with an average of 8.5 alleles per locus. When tested on wild perch from a population in Pennsylvania, observed and expected heterozygosities ranged from 0.07-0.81 and from 0.37-0.95, respectively. Of 2,226 EST sequences examined, only 110 (4.93%) contained microsatellites and for those, 13 markers were tested, 12 of which exhibited polymorphism. Compared with genomic-SSRs, EST-SSRs exhibited a lower level of genetic variability with the number of alleles of averaging only 2.6 alleles per locus. Cross-species utility indicated that three of the genomic-SSRs and eight of the EST-SSRs successfully cross-amplified in a related species, the walleye (Sander vitreus).