To identify the effects of the expression levels of DNMT3a and DNMT3b, coding the de novo methyltransferases DNMT3a and DNMT3b, on 16 beef carcass and quality traits, 50 beef cattle liver and ribeye muscle tissue samples were collected. Quantitative real-time RT-PCR was employed to quantify the expression level of these two genes, and a basic model included fixed effects of gender, age, and expression level of these two genes was used to analyze live weight; and slaughtering batches and aging days were added when beef carcass traits and beef quality traits were analyzed, respectively. Results showed that transcripts of DNMT3a and DNMT3b were present at significantly higher levels in liver tissue than in muscle tissue, and the expression level of DNMT3a was significantly higher than that of DNMT3b in both tissues. Regression analysis found that the expression levels of DNMT3a and DNMT3b were associated with several beef quality traits, which are important in beef breeding. Findings of the present study suggested that these two genes could significantly contribute to the improvement of beef quality genetically.