No-tillage (NT) farming is superior to intensive tillage for conserving soil and water, yet its potential for sequestering soil organic carbon (SOC) in all environments as well as its impacts on soil profile SOC distribution are not well understood. Thus, we assessed the impacts of long-term NT-based cropping systems on SOC sequestration for the whole soil profile (0–60-cm soil depth) across 11 Major Land Resource Areas (MLRAs: 121, 122, and 125 in Kentucky; 99, 124, 139A in Ohio; and 139B, 139C, 140, 147, and 148 in Pennsylvania) in the eastern United States. Soil was sampled in paired NT and plow tillage (PT) based cropping systems and an adjacent woodlot (WL). No-tillage farming impacts on SOC and N were soil specific. The SOC and N concentrations in NT soils were greater than those in PT soils in 5 out of 11 MLRAs (121, 122, 124, 139A, and 148), but only within the 0- to 10-cm depth. Below 10 cm, NT soils had lower SOC than PT soils in MLRA 124. The total SOC with NT for the whole soil profile (0–60 cm) did not differ from that with PT (P > 0.10) in accord with several previous studies. In fact, total soil profile SOC in PT soils was 50% higher in MLRA 125, 21% in MLRA 99, and 41% in MLRA 124 compared with that in NT soils. Overall, this study shows that NT farming increases SOC concentrations in the upper layers of some soils, but it does not store SOC more than PT soils for the whole soil profile.