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Fig. 1 | Genome Biology

Fig. 1

From: NT-seq: a chemical-based sequencing method for genomic methylome profiling

Fig. 1

Principle and workflow of NT-seq. a Schematic illustration of nitrite treatment. Nitrite treatment induces deamination of adenine, cytosine, and 5mC at different frequencies, producing inosine, uracil, and thymine, respectively. Meanwhile, nitrite treatment nitrosylates 6mA and 4mC, producing nitrosylated 6mA (6mA-NO) and nitrosylated 4mC (4mC-NO). During PCR amplification and sequencing, base pairing and reading for each product are labeled on the right column. b The workflow of NT-seq. Single-stranded DNA is first annealed with protective oligos to protect PCR primer regions. Annealed DNA is treated with nitrite and then amplified to construct the sequencing library. Sequencing data from native DNA and PCR control are used to calculate the A to G or C to T mutation ratio and to call methylation

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