title: A novel combined bisulfite UDG assay for selective 5-methylcytosine detection.
abstract: DNA modification, a significant epigenetic event, largely affects genes' binding with the transcription factors and some other DNA binding proteins. Among DNA modifications, methylation, especially cytosine methylation is of great importance and attracts extensive studies as it leads to the silence of tumor-suppressor gene expression. In this work, a novel combined bisulfite Uracil-DNA glycosylase (UDG) assay has been developed on the basis of bisulfite modification to generate uracil from cytosine, subsequent UDG-mediated uracil elimination and ultimate DNA cleavage in alkaline condition. This strategy can be used to selectively detect exact number and loci of 5-methylcytosine residues regardless of sequence context. Moreover, it provides linear quantitative results of DNA methylation level across a wide range.
authors.affiliation: College of Chemistry and Molecular Sciences, Key Laboratory of Biomedical Polymers of Ministry of Education, Wuhan University, Hubei, Wuhan 430072, PR China.
authors.forename: Rong
authors.initials: R
authors.lastname: Huang
authors.forename: Jiaqi
authors.initials: J
authors.lastname: Wang
authors.forename: Wuxiang
authors.initials: W
authors.lastname: Mao
authors.forename: Boshi
authors.initials: B
authors.lastname: Fu
authors.forename: Xiwen
authors.initials: X
authors.lastname: Xing
authors.forename: Ge
authors.initials: G
authors.lastname: Guo
authors.forename: Xiang
authors.initials: X
authors.lastname: Zhou
mesh_terms: D044503:5-Methylcytosine / Q000032:analysis* / Q000737:chemistry; D003596:Cytosine / Q000737:chemistry; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D053837:DNA Cleavage; D019175:DNA Methylation; D004591:Electrophoresis, Polyacrylamide Gel; D057075:Enzyme Assays; D006801:Humans; D009838:Oligodeoxyribonucleotides / Q000032:analysis* / Q000737:chemistry; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012972:Sodium Hydroxide; D012996:Solutions; D013447:Sulfites / Q000737:chemistry; D014498:Uracil / Q000032:analysis* / Q000138:chemical synthesis; D051981:Uracil-DNA Glycosidase / Q000737:chemistry*
chemical_list: D009838:Oligodeoxyribonucleotides; D012996:Solutions; D013447:Sulfites; D012972:Sodium Hydroxide; D014498:Uracil; D044503:5-Methylcytosine; D003596:Cytosine; D004247:DNA; D051981:Uracil-DNA Glycosidase; C009279:sodium bisulfite
keywords: Bisulfite modification; Methylation; Methylcytosine; PAGE; Uracil DNA glucosylase