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Fig. 1 | BMC Genomics

Fig. 1

From: Universal probe-based intermediate primer-triggered qPCR (UPIP-qPCR) for SNP genotyping

Fig. 1

Principle and application of UPIP-qPCR for SNP genotyping. The reactions of UPIP-qPCR are divided into two stages (a). The first stage is a general PCR reaction aiming to obtain a certain amount of DNA fragments containing specific SNP sites (a, upper panel in each sample). The second stage is a quantitative PCR (qPCR) reaction, which uses the PCR products of the first stage as templates to obtain the corresponding fluorescence signals of alleles (a, lower panel in each sample). UPIP-qPCR was initially used to identifying the SNP genotypes of ALDH2 rs671 (b). All three genotypes of each SNP samples successfully yielded accurate results by UPIP-qPCR. The homozygous WT genotypes presented by FAM-only signals (blue curves) were shown in the lower right area of the genotyping scatter diagrams with yellow dots, the heterozygous genotypes presented by FAM and HEX signals (blue curves and green curves) were shown in the middle area of the genotyping scatter diagrams with green triangles, and the homozygous mutant genotypes presented by HEX-only signals (green curves) were shown in the upper left area of the genotyping scatter diagrams with blue squares. No-template control groups (NTC) were shown in the lower left area of the genotyping scatter diagrams with black rhombuses. Each sample was detected with three duplicates in one experiment, and the experiments were repeated more than three times

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