Tissue-Specific Methylation of Human Insulin Gene and PCR Assay for Monitoring Beta Cell Death
Figure 4
Primer selection and analytical performance of methylation-specific PCR.
A) Schematic illustration of the human INS gene promoter region showing the position of the nine CpG sites. Solid arrows represent the bisulfite-specific primers (BSPs) that amplify both methylated and unmethylated DNA. Dashed arrows represent methylation-specific primers (MSPs) that amplify unmethylated DNA only. B) Unmethylated plasmid was serially diluted after bisulfite conversion and analyzed by qMSP using selected primer sets. Agarose gel electrophoresis of MSP reactions showing the size of the PCR products. C) Graphs of real-time SYBR Green PCR data showing linearity of Cq versus log copy number of unmethylated plasmid (averages and standard deviation (SD)) from 5 to 106 copies.