Mirror Bisulfite Sequencing: A Method for Single-Base Resolution of Hydroxymethylcytosine.
Anal Chem
; 90(22): 13200-13206, 2018 11 20.
Article
em En
| MEDLINE
| ID: mdl-30345735
ABSTRACT
Although the role of 5-methylcytosine has been well studied, the biological role of 5-hydroxymethylcytosine still remains unclear because of the limited methods available for single-base detection of 5-hydroxymethylcytosine (5hmC). Here, we present mirror bisulfite sequencing for 5hmC detection at a single CpG site by synthesizing a DNA strand to mirror the parental strand. This semiconservative duplex is sequentially treated with ß-glucosyltransferase and M.SssI methylase. The 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 genomewide.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Sulfitos
/
DNA
/
Análise de Sequência de DNA
/
5-Metilcitosina
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
En
Ano de publicação:
2018
Tipo de documento:
Article