[Analysis Case] SNP Analysis Using Real-Time PCR Method
SNP analysis is possible using real-time PCR (probe method).
Real-time PCR is a method that detects the amplification of DNA fragments from PCR reactions as a fluorescent signal for each cycle. This introduces SNP (Single Nucleotide Polymorphism) analysis using the probe method of real-time PCR. Specific base sequences (probes) labeled with a reporter (fluorescent dye) and a quencher (quenching substance) are used to repeat the three steps shown below: 1) to 3). By using different reporters for each base, the SNP base is identified from the fluorescence intensity. Reactions at each step: 1) Thermal denaturation DNA is converted from double-stranded to single-stranded. 2) Annealing and hybridization Primers and probes specifically bind to single-stranded DNA. 3) Extension reaction The probe is degraded by the extension reaction of the primer, releasing the reporter from the quencher, resulting in fluorescence.
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Analysis of biotechnology, pharmaceuticals, cosmetics, food, and the environment.