PT  - JOURNAL ARTICLE
AU  - Tan, Darany
AU  - Chung, Tzu Hung
AU  - Sun, Xueguang
AU  - Jia, Xi-Yu
TI  - Mirror bisulfite sequencing — a method for single-base resolution of hydroxymethylcytosine
AID  - 10.1101/019943
DP  - 2015 Jan 01
TA  - bioRxiv
PG  - 019943
4099  - http://biorxiv.org/content/early/2015/05/29/019943.short
4100  - http://biorxiv.org/content/early/2015/05/29/019943.full
AB  - 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 β- 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.