A rapid, quantitative, non-radioactive bisulfite-SNuPE-IP RP HPLC assay for methylation analysis at specific CpG sites

被引：57

作者：

El-Maarri, O

Herbiniaux, U

Walter, J

Oldenburg, J

机构：

[1] Inst Expt Haematol & Transfus Med, D-53105 Bonn, Germany

[2] Univ Saarland, D-66041 Saarbrucken, Germany

[3] Inst Transfus Med & Immune Haematol, D-60528 Frankfurt, Germany

来源：

NUCLEIC ACIDS RESEARCH | 2002年 / 30卷 / 06期

关键词：

D O I：

10.1093/nar/30.6.e25

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The precise mapping and quantification of DNA methylation as an epigenetic parameter during development and in diseased tissues is of great importance for functional genomics. Here we describe a rapid, quantitative method to assess methylation levels at specific CpG sites using PCR products of bisulfite-treated genomic DNA. Using single nucleotide primer extension (SNuPE) assays in combination with Ion pair reverse phase high performance liquid chromatography (IP RP HPLC) separation techniques, methylated and unmethylated CpGs can be discriminated and quantified based on the different masses and hydrophobicities of the extended primer products. The assay is linear, highly reproducible and several sites can be measured simultaneously in one reaction. It can be semi-automated and eliminates the need for cloning and sequencing of individual bisulfite PCR products.

引用

页数：4

共 8 条

[1] Methylation levels at selected CpG sites in the factor VIII and FGFR3 genes, in mature female and male germ cells: Implications for male-driven evolution [J].