Biodiesel consisting principally of monounsaturated fatty acid methyl esters (FAME) has been reported to have the optimal balance between cold flow properties and oxidative stability, therefore producing a superior fuel. In addition, treating biodiesel with antioxidants such as selenium (Se) also increases oxidative stability. Fuel properties including acid value (AV), cloud point (CP), iodine value (IV), pour point (PP), peroxide value (PV), induction period (IP), onset temperature (OT), and kinematic viscosity (KV) were used to evaluate a newly developed Roundup Ready® soybean recombinant inbred line (RIL) and a commercial cultivar. The RIL had a fatty acid profile with elevated levels of monounsaturated FAME. TN07‐93RR was determined as the more desirable line for production of biodiesel, based on its fatty acid composition and subsequent fuel properties. The commercial cultivar AG3906 contained the highest abundance of polyunsaturated FAME and exhibited comparatively high IV and low oxidative stability. AG3906 was therefore not acceptable under the European biodiesel standard, EN 14214. However, TN07‐93RR and AG3906 both were considered satisfactory according to the American biodiesel standard, ASTM D6751. Foliar treatment of soybean plants with varying amounts of Se had no effect on subsequent biodiesel oxidative stabilities.