Continuous no-till (NT) can be beneficial relative to tillage with fewer field operations, reduced erosion, and surface soil improvement. Field research was conducted at two locations for 5 yr in eastern Nebraska to test the hypotheses that one-time tillage of NT can result in increased grain yield, reduced stratification of soil properties persisting for at least 5 yr, a net gain in soil organic carbon (SOC), and a restoration of the soil microbial community to NT composition. Stratification of soil test P, SOC, and bulk density was similar for all tillage treatments at 5 yr after tillage. Water stable soil aggregates (WSA) were not affected by tillage treatments except that there was more soil as macroaggregates at one location in the 5- to 10-cm depth with moldboard plow tillage (MP) compared with NT. Tillage treatments had no effect on SOC mass in the 0- to 30-cm depth. Soil microbial biomass was greater at the 0- to 5-cm compared with the 5- to 10-cm depth. Biomass of bacteria, actinomycetes, and arbuscular mycorrhizal fungi was greater with NT compared with one-time MP at one location but not affected by the one-time tillage at the other location. Microbial community structure differed among tillage treatments at the 0- to 5-cm depth at one location but not at the other location. Grain yield generally was not affected by tillage treatment. One-time tillage of NT can be done without measureable effects on yield or soil properties.