Glyphosate has been demonstrated to reduce the macronutrient and micronutrient content of glyphosate-susceptible (GS) and first generation glyphosate-resistant (GR) or Roundup Ready (RR) soybean, possibly by complexation of the herbicide molecule with the nutrient. The recent release of newer GR soybean cultivars, second generation Roundup Ready 2 Yield (RR2Y), provides growers with newer technology for weed management programs, but it is unclear how the nutrient content of these cultivars is affected by glyphosate in a field setting. The objective of this experiment was to identify the effect of glyphosate on the concentration of macronutrient and micronutrients in RR and RR2Y soybean when grown using standard agronomic practices in Indiana. The macronutrients analyzed were nitrogen, phosphorus, potassium, sulphur, magnesium, and calcium. The micronutrients analyzed were boron, zinc, manganese, iron, copper, and aluminum. Our results indicate that while differences in accumulation of macro and micronutrients exist between the two cultivars tested, there was no consistent effect due to glyphosate treatment. Glyphosate-induced deficiency symptoms observed in previous reports were not observed in this study. Growers should continue to monitor soil nutrient levels to identify and correct nutrient deficiencies.