GuideDiagnostic Laboratory Methods

Bisulfite Sequencing

A DNA sequencing technique that can differentiate cytosine from 5-methylcytosine in a DNA sample. First, a denatured DNA sample is treated with bisulfite which converts non-methylated cytosine to uracil. Next, the sample is amplified using a PCR method that does not discriminate between non-methylated and methylated sequences. The amplified DNA is subjected to nucleotide sequencing. The resulting sequence is compared to an identical control sample of DNA that was not treated with bisulfite. Unmethylated cytosines will be displayed as cytosines in the control sample and as thymines in the bisulfite-treated sample.

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Updated
2026-10-18
Author
National Cancer Institute (source definition)
Reviewer
Automated source check (NCI definition)

What it is

A DNA sequencing technique that can differentiate cytosine from 5-methylcytosine in a DNA sample. First, a denatured DNA sample is treated with bisulfite which converts non-methylated cytosine to uracil. Next, the sample is amplified using a PCR method that does not discriminate between non-methylated and methylated sequences. The amplified DNA is subjected to nucleotide sequencing. The resulting sequence is compared to an identical control sample of DNA that was not treated with bisulfite. Unmethylated cytosines will be displayed as cytosines in the control sample and as thymines in the bisulfite-treated sample.

Source: National Cancer Institute, NCI Thesaurus 26.09d, C106054. NCI definition, reproduced without modification under CC BY 4.0.

Source identity and definition checked automatically on 2026-10-18. No individual clinical interpretation or human reviewer is asserted.

Compliance note

Reference intervals and interpretation depend on the laboratory, specimen, method and clinical context. This source definition is educational information; it does not establish a diagnosis, product approval or suitability for an individual.

Bisulfite Sequencing | IVDmap