Abstract: In this experiment, specific PCR primers and Taqman probes were designed based on the differential mitochondrial DNA sequences of bovine mitochondria, and based on the 16S rDNA internal reference gene, universal primers and Taqman probes were designed. The total DNA concentration and DNA concentration of bovine source samples were determined by standard curve. Relative quantification method was used to determine the quality fraction of bovine source in meat products. The method was validated by specificity, sensitivity experiments, mixed meat recovery, and commercial meat testing. The results show that that method is specific and can amplify specific DNA of beef. The minimum observable limit with high sensitivity is 10 pg/μL.According to the recovery test, the average recovery was 98.62%, which can provide technical reference for the determination of beef content in meat products.