Soybean seeds contain high amounts of isoflavones that display biological effects and isoflavone content of soybean seed can vary by year, environment, and genotype. Objective of this study was to identify quantitative trait loci that underlie isoflavone content in soybean seeds. The study involved 85 F2 populations derived from Korean soybean cultivar 'Kwangkyo' and wild type soybean 'IT182305' for QTL analysis associated with isoflavone content. Isoflavone content of seeds was determined by HPLC. The genetic map of 33 linkage groups with 207 markers was constructed. The linkage map spanned 2,607.5 cM across all 33 linkage groups. The average linkage distance between pair of markers among all linkage groups was 12.6 cM in Kosambi map units. Isoflavone content in F2 generations varied in a fashion that suggested a continuous, polygenic inheritance. Eleven markers (4 RAPD, 3 SSR, 4 AFLP) were significantly associated with isoflavone content. Only two markers, Satt419 and CTCGAG3 had F-tests that were significant at P<0.01 in F2 generation for isoflavone content. Interval mapping using the F2 data revealed only two putative QTLs for isoflavone content. The peak QTL region on linkage group 3, which was near OPAG03c, explained 14~% variation for isoflavone content. The peak QTL region on linkage group 5, which was located near OPN14 accounted for 35.3~% variation for isoflavone content. Using both Map-Maker-QTL (LOD~geq2.0) and single-factor analysis (P~leq0.05) , one marker, CTCGAG3 in linkage group 3 was associated with QTLs for isoflavone content. This information would then be used in identification of QTLs for isoflavone content with precision