RT Journal Article
SR Electronic
T1 Mirror bisulfite sequencing — a method for single-base resolution of hydroxymethylcytosine
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 019943
DO 10.1101/019943
A1 Darany Tan
A1 Tzu Hung Chung
A1 Xueguang Sun
A1 Xi-Yu Jia
YR 2015
UL http://biorxiv.org/content/early/2015/05/29/019943.abstract
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.