A total of 45 sows (Landrace × Yorkshire) and their litters were used in this study to evaluate the efficacy of nucleotide supplementation on reproductive performance, growth performance, fecal microflora, and blood profiles in sows and piglets. Sows were allocated to one of three treatments (15 pigs per treatment and the average parity was 4.13). Dietary treatments consisted of (1) CON: basal diet, (2) T1: CON + 0.5% nucleotides, and (3) T2: CON + 1.0% nucleotides. In this study, the average daily feed intake during lactation and back fat at weaning day was increased linearly (P < 0.05) in sows with the increase in nucleotide supplementation levels. Sows receiving increasing levels of nucleotides had linearly increased fecal Lactobacillus counts and decreased Escherichia coli counts at weaning day (P < 0.05). Linear reduction of the epinephrine, norepinephrine, and cortisol in sows was observed at weaning day, whereas superoxide dismutase was increased linearly (P < 0.05). Body weight and average daily gain in piglets were linearly (P < 0.05) increased with the increasing levels of nucleotides. Linear increase (P < 0.05) was also observed on total piglets born, live piglets, and piglet survival. The current study demonstrated the importance of nucleotides in the diets of sows to improve sow and piglet performance.