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bioRxiv · 10.1101/019943

Mirror bisulfite sequencing — a method for single-base resolution of hydroxymethylcytosine

Xueguang Sun·Darany Tan·Tzu Hung Chung·Xi-Yu Jia
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Abstract

While the role of 5-methylcytosine has been well studied, the biological role of 5- hydroxymethylcytosine still remains unclear due to the limited methods available for single-base detection of 5-hydroxymethylcytosine (5hmC). Here, we present Mirror bisulfite sequencing detects 5-hydroxymethylcytosines at a single CpG site by synthesizing a DNA strand to mirror the parental strand. This semi-conservative duplex is sequentially treated with {beta}- glucosyltransferase and M.SssI methylase. A glucosyl-5hmCpG in the parental strand inhibits methylation of the mirroring CpG site, and after bisulfite conversion, a thymine in the mirroring strand indicates a 5hmCpG site in the parental strand whereas a cytosine indicates a non-5hmC site. Using this method, the 5hmC levels of various human tissues and paired liver tissues were mapped genome-wide.

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Xueguang Sun, Darany Tan, Tzu Hung Chung, Xi-Yu Jia. 2015-05-29. Mirror bisulfite sequencing — a method for single-base resolution of hydroxymethylcytosine. https://doi.org/10.1101/019943

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