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Schematic diagram of direct bisulfite sequencing workflow. DNA is first bisulfite treated to conserve methylated cytosine while unmethylated cytosine is converted to uracil. During PCR, uracil is converted to thymine and product is amplified. PCR product is then sequenced. The methylation level of each CpG in the amplicon will result in differing peak sizes for cytosine and thymine as read on the chromatogram.
