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Schematic of bisulfite PCR–ligase detection reaction–real-time quantitative PCR assay for detection of CpG methylation. 1. Bisulfite conversion. 2. Linear amplification (reverse primer only). 3. Digestion by uracil-N-DNA glycosylase (UDG). 4. PCR (add forward primer). 5. Ligase detection reaction. 6. Digestion by UDG. 7. ViiA7 real-time PCR.
