The influence of soaking time in deep cryogenic treatment on the tensile and impact properties of low-alloy medium-carbon HY-TUF steel was investigated in this study. Microstructural studies based on phase distribution mapping by electron backscatter diffraction show that the deep cryogenic process causes a decrease in the content of retained austenite and an increase in the volume fraction of η-carbide with increasing soaking time up to 48 h. The decrease in the content of retained austenite from ~1.23vol% to 0.48vol% suggests an isothermal martensitic transformation at 77 K. The η-type precipitates formed in deep cryogenic-treated martensite over 48 h have the Hirotsu and Nagakura orientation relation with the martensitic matrix. Furthermore, a high coherency between η-carbide and the martensitic matrix is observed by high-resolution transmission electron microscopy. The variations in macrohardness, yield strength, ultimate tensile strength, and ductility with soaking time in the deep cryogenic process show a peak/plateau trend.