通过简化代表性亚硫酸氢盐测序对低细胞数样本进行全基因组DNA甲基化分析。

Genome-wide analysis of DNA methylation in low cell numbers by reduced representation bisulfite sequencing.

作者信息

Smallwood Sébastien A, Kelsey Gavin

机构信息

Epigenetics Programme, The Babraham Institute, Cambridge, UK.

出版信息

Methods Mol Biol. 2012;925:187-97. doi: 10.1007/978-1-62703-011-3_12.

DOI:10.1007/978-1-62703-011-3_12

PMID:22907498

Abstract

Development of high-throughput sequencing technologies now enables genome-wide analysis of DNA methylation of mammalian cells and tissues. Here, we present a protocol for Reduced Representation Bisulfite Sequencing (RRBS) applicable to low amounts of starting material (from 200 to 5,000 cells). RRBS is a cost-effective and powerful technique offering the advantages of absolute DNA methylation quantification and single nucleotide resolution while covering mainly CpG islands. Typically one sequencing experiment using the Illumina Genome Analyser IIx platform provides information on the DNA methylation status of more than half of the CpG islands of the mouse genome.

摘要

高通量测序技术的发展使得对哺乳动物细胞和组织的DNA甲基化进行全基因组分析成为可能。在此，我们介绍一种适用于少量起始材料（200至5000个细胞）的简化代表性亚硫酸氢盐测序（RRBS）方案。RRBS是一种经济高效且强大的技术，具有绝对DNA甲基化定量和单核苷酸分辨率的优势，同时主要覆盖CpG岛。通常，使用Illumina Genome Analyser IIx平台进行一次测序实验就能提供有关小鼠基因组一半以上CpG岛DNA甲基化状态的信息。