Broiler litter (BL) is a high value manure available to Kentucky crop producers to utilize as a fertilizer. The rate of BL application, timing of application, and method of application are all important factors to take into account when utilizing BL. A laboratory incubation was conducted to observe nitrogen (N) mineralization rates of BL amended soils over time under different conditions. Different application methods, application rates, watering frequencies, and bedding materials were used to determine their influence on N mineralization. Broiler litter applied on the soil surface, at lower rates, at lower watering frequency resulted in lower mineralization rates than BL that was incorporated, at higher rates and higher watering frequency. The rice hull bedding BL treatments had a significantly higher initial amount of NH4+-N than the wood-based bedding materials. The increased NH4+-N content produced a lower pH due to higher rates of nitrification. The difference in NH4+-N resulted in the rice hull BL treatments containing more inorganic N throughout the incubation. The wood-based BL contained significantly more carbon (C), zinc (Zn), and potassium (K). The results collected will help improve current nutrient recommendations and provide information that will help utilize BL more efficiently and economically.