Nitrogen (N) is the most significant nutrient affecting crop growth and development for all types of crops, except legumes. The goal of this study was to optimize the N level for cotton grown in a semi-arid environment to enhance growth and development, determine N status, and increase seed cotton yield and biomass. Two independent field experiments each three years in duration were conducted, from 2007 to 2009 (Exp.-I) and 2018 to 2020 (Exp.-II). Experiments were laid out in a randomized complete block design with three replicates. The N treatments in Exp.-I were comprised of 0, 40, 80, 120, 160, 200 and 240 kg N/ha, while treatments in Exp.-II were comprised of 0, 70, 140, 210 and 280 kg N/ha. A wide range of data sets for cotton traits were recorded, including canopy height, leaf area index, the N status of the leaf and stem, seed cotton yield and time-series biomass data. The higher N rates 240 and 280 kg N/ha performed better for all these traits. However, the highest leaf N contents were recorded for 210 kg N/ha. Based on these results, it is suggested that under semi-arid conditions, slightly higher rates than optimum or recommended N rates could be applied as a strategy by cotton growers for a higher seed cotton yield. The findings of this study may also increase profitability in other cotton-growing areas that have similar weather conditions.