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To ascertain an appropriate level of isoleucine for LSL-LITE layers (23- to 30-week-old), diets containing total isoleucine concentrations (levels) of 0.66 (Control), 0.69, 0.72, 0.75, 0.78, 0.81, and 0.84% were fed as 7 treatments (2730 kcal/kg metabolizable energy) x 7 replicates x 10 birds per replicate. Significance for performance, egg quality, serum biochemistry, and ileal digestibility of protein was determined at P ≤ 0.05. Level, week, and level*week (L*W) were significant for production, egg mass, and feed intake. Level and week were significant for FCR. Week was significant for weight gain. Level was significant for egg weight, specific gravity, and shell thickness; week was also significant for these external egg parameters as well as shape index and proportional shell thickness. L*W was significant for all except shape index. For internal egg measurements, level was significant for proportional yolk, proportional albumen, yolk index, and yolk:albumen. Week was significant for internal egg parameters while L*W significantly affected Haugh unit, proportional albumen weight, yolk index, albumen index, and yolk color. Level was significant for globulin and glucose in serum. Isoleucine at 0.72%, 0.81%, and 0.84% produced the lowest FCR, an important standard in the poultry industry. Considering the low FCR of 1.45 and cost for inclusion as a dietary ingredient, 0.72% isoleucine was chosen for further studies with varying quantities of other branched chain amino acids in diets for young laying hens.