Traditional differentiation of myoblasts is usually induced by serum starvation method, in which 2%HS is used to induce myoblasts to differentiate into myotubes. But, traditional serum starvation method is difficult to differentiate large yellow croaker muscle satellite cells efficiently. Although myotubes were induced within 2-3 days with medium containing 2% horse serum (HS), they detached shortly and died soon after. To improve differentiation efficiency and survival rate, different combinations of basal medium, serum ratios and myogenic factors were evaluated. Finally, the F12 medium containing 8% HS, 10 ng/ml IGF-1, 50 nM necrosulfonamide and 200 µM ascorbic acid was identified as an effective recipe for myogenic differentiation. On day 3 of culture in this differentiation medium, elongated myotubes began to appear, and on day 6, striations similar to skeletal muscle were observed in some of myotubes. But it is still inefficient, the fusion index (the proportion of nuclei in multinucleated myotubes) was 6.39% on day 3 and 1.30% on day 6 (1.30%) .Therefore, we need to find a more efficient method to induce myogenic differentiation. We then performed gene expression profiling analysis using data obtained from RNA-seq of Large yellow croaker muscle satellite cells during differentiation at three time points(day0,day3,day6). Overall design: differential expression analysis of RNA-seq data for Large yellow croaker muscle satellite cells during differentiation at three time points(day0,day3,day6)