One of the alternative sources for petroleum is soybean biodiesel. Developing high oil soybeans would reduce the cost of biodiesel production. The objectives of this study were to: develop a single seed oil measurement method using NMR spectrometryï¼› evaluate the potential of increasing the oil content and determine the genetic basis of the trait. Five crosses were made between RCAT Angora, OAC Wallace, and OAC Glencoe. Single seed NMR method for oil measurement was developed. Seventy oil related SSR markers were evaluated in two F 2 populations. Transgressive segregation was observed with oil content ranging between 16.3 and 27.0% at the F2 and, 12.2 and 25.7% at the F3 generations. Additive genetic effects ranged between 4.09% and 61.84%, dominance between 2.45% and 73.62%, and epistasis between 0% and 49.6%. Best fit linear regression models including individual oil QTL and epistasis explained up to 75.8% of variation per population.