Primer pair and probe for detecting sheep-derived component, kit and application thereof
A sheep-derived, kit-based technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of restricting the application of food and meat detection, lengthy and other problems, and achieve simple and fast operation, Accurate results and high detection sensitivity
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[0066] 1. Sample collection
[0067] Use sterile scissors to save the sample to be tested into a sterile centrifuge tube for later use.
[0068] 2. DNA extraction
[0069] The DNA of the meat samples to be tested was extracted with 5% Chelex-100.
[0070] 3. RAA-LFD
[0071] 3.1. Design of primers and probes for specific sequence amplification of sheep-derived components
[0072] Download the complete mitochondrial genome sequence of sheep (NC_001941.1), use the sequence of the sheep D-loop gene as the target gene, analyze the characteristics of the sheep sequence and compare it with the sequences of common adulterated meat species, including duck (NC_009684.1), Cattle (NC_006853.1), pig (NC_000845.1), chicken (NC_040902.1), species-specific amplification primers were designed using Primer Premier 5.0 software. The probe sequence is located in the middle section of the amplification primer and modified by tetrahydrofuran. Primer design follows the following principles:
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