Promoter sequence of gene MyoG in duck was amplified and cloned using RT–PCR, and its bioinformatic info was analyzed as well; the methylation level in CpG island (from –2 536 to –1 997 bp) of promoter MyoG in muscle tissues was detected using the Sequenom MassArray technique, meanwhile, its expression level was detected using qRT–PCR. The results showed that the amplified MyoG promoter sequences was 2 730 bp which contained 2 CpG islands in promoter region. The CpG island (from –2 536 to –1 997 bp) included 5 binding sites of transcription factor and several eukaryotes structure components. The methylation data showed that methylation status from different individuals and tissues did not clustered together. Methylation frequency in CpG loci varied with individuals, and the methylation frequency on 22% of locus were negatively correlated with MyoG expression levels (P>0.05), whereas, on 78% of locus were positively correlated (P>0.05) with that, among of them, they reached significant difference level on locus of CpG 1, CpG 26.27.28.29 from leg muscles (P<0.05). The mechanism of regulating transcription for MyoG was different from mammals. Among the potential locus influencing MyoG transcription, CpG_1 and CpG_26.27.28.29 might affect MyoG transcription in skeletal muscle tissues of duck leg through DNA methylation. This study might list foundation data for studying the regulatory mechanisms of MyoG transcription in duck.