Real- time PCR detection of sheep derived materials in meat product

Targeting cytochrome b gene of mitochondrial DNA from sheep,the specific primers and Taq Man probes were designed.After optimization of reaction conditions and establishment of standard curve,a fluorescent realtime PCR method had been set up to detect sheep derived materials.The results showed that the specific primers could identify sheep derived materials in meat product specially and effectively,and the sensitivity was as high as16 pg / μL.Amplification efficiency of the standard curve was 98.561% and correlation coefficient R2 was 1.000.These properties meet the requirement of the《MIQE Guide》.The results obtained according to the new method and the National Standard( GB / T20190Y-2006 routine PCR method) were exactly the same,and electrophoresis,restriction digestion and sequencing were avoided in the new method. The value of this method was proved though the identification of the meat product from the market.The assay provides a new way for the quality control of the meat products.