Identification of Dangxiong Alpine Yak Meat by Single Nucleotide Polymorphism-Based Multiplex Real-Time Fluorescent Quantitative Polymerase Chain Reaction
LIU Ruixi, QIMEICIREN, LI Jiapeng, WEI Yixuan, LI Jinchun, WANG Shouwei
1.Beijing Key Laboratory of Meat Processing Technology, China Meat Food Research Center, Beijing Academy of Food Science, Beijing 100068, China; 2.The People’s Government of Dangxiong County, Tibet, Lhasa 851500, China
Abstract：By comparing the difference in the mitochondrial gene of yaks from five different regions, 11 single nucleotide polymorphism (SNP) loci were selected for identification of the meat of Dangxiong alpine yaks. Utilizing the SNP loci as the 3’ end of primers, and introducing the concept of base mismatch, 8 pairs of specific primers were designed to construct two triplex and one duplex real time polymerase chain reaction (RT-PCR) reaction systems. An SNP-based multiplex RT-PCR method was finally established to identify the meat of Dangxiong alpine yaks. The melting temperatures (Tm) of the three RT-PCR reaction systems were reaction A: 73.1–75.2, 76.0–77.3, and 79.0–80.0 ℃; reaction B: 69.5–72.8, 75.0–77.9, and 78.5–80.0 ℃; and reaction C: 76.1–78.1, and 79.5–80.7 ℃, respectively. The determination of whether an unknown sample was the meat of Dangxiong alpine yaks was dependent on the presence or absence of melting curve peaks in the three reaction systems and whether the peaks corresponded to the Tm ranges. The method was successfully applied to the rapid identification of real samples .
2020, Vol. 34 
