Figure 1 | Scientific Reports

Figure 1

From: Quantification of allelic differential expression using a simple Fluorescence primer PCR-RFLP-based method

Figure 1

Schematic diagram of Fluorescence PCR-RFLP (fPCR-RFLP) method. Total RNA was extracted from tissues or cells. cDNAs were synthesized by RT-PCR. Heteroduplex was validated by genotyping and then subjected to the first round of PCR (about 35 cycles) using a gene-specific primer. Next, the PCR product was used as template for the second round of PCR with one cycle using one fluorescently labeled primer. This step generated two alleles with over-hang labeling. The labeled PCR products were digested by restriction enzyme. The digested products were separated by agarose gel electrophoresis. Finally, only the fluorescently labeled fragments were detected by a laser detection system. The ratio of peak area represented the ADE.

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