Bisulfite sequencing infers cytosine modification from differential chemical conversion before DNA sequencing. Under the relevant chemistry, unmodified cytosine is converted to uracil and is subsequently read as thymine, while 5-methylcytosine is retained as cytosine. Incomplete conversion, sequence coverage and strand assignment affect interpretation. Conventional bisulfite sequencing does not by itself separate 5-methylcytosine from every other conversion-resistant modification, so its signal requires a stated chemical scope. [ref]

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Bisulfite sequencing is a method used to determine the methylation status of DNA, particularly focusing on the conversion of cytosine residues in DNA. This technique exploits the unique properties of sodium bisulfite, which converts unmethylated cytosines into uracils (which are subsequently treated as thymidines during PCR amplification), while leaving methylated cytosines unchanged.
Bisulfite sequencing by Ciro Santilli 40 Updated 2025-07-16
The main way to sequence DNA methylation. Converts methylated cytosine to uracil, and then we can sequence those.
Video 1.
Bisulfite Sequencing by Henrik's Lab (2020)
Source. Nothing much new that we don't understand from a single sentence in the animation. But hey, animations!