Quantitative methylation analysis using methylation-sensitive single-nucleotide primer extension (Ms-SNuPE)

被引：49

作者：

Gonzalgo, ML ^{[1
]}

Jones, PA ^{[1
]}

机构：

[1] Univ So Calif, Kenneth Norris Jr Comprehens Canc Ctr, Keck Sch Med, Dept Biochem & Mol Biol,Urol Canc Res Lab, Los Angeles, CA 90089 USA

来源：

METHODS | 2002年 / 27卷 / 02期

关键词：

D O I：

10.1016/S1046-2023(02)00064-6

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Methylation-sensitive single-nucleotide primer extension (Ms-SNuPE) is a technique that allows for rapid and simultaneous quantitation of the degree of methylation at several CpG sites. Treatment of genomic DNA with sodium bisulfite is used to convert unmethylated cytosine to uracil while leaving 5-methylcytosine unchanged. A strand-specific polymerase chain reaction product is then generated to provide a suitable DNA template for quantitative methylation analysis using Ms-SNuPE. Single-nucleotide primer extension is performed with oligonucleotide(s) designed to hybridize immediately upstream of the CpG site(s) being analyzed. The Ms-SNuPE technique can be adapted for high-throughput methylation analysis and therefore represents a novel approach for rapid quantitation of cytosine methylation suitable for a wide range of biological investigations. (C) 2002 Elsevier Science (USA). All rights reserved.

引用

页码：128 / 133

页数：6

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