用于从脑组织中以基因和核苷酸分辨率检测DNA甲基化的直接亚硫酸氢盐测序。
Direct bisulfite sequencing for examination of DNA methylation with gene and nucleotide resolution from brain tissues.
作者信息
Parrish R Ryley, Day Jeremy J, Lubin Farah D
机构信息
Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of Alabama at Birmingham, Birmingham, Alabama, USA.
出版信息
Curr Protoc Neurosci. 2012 Jul;Chapter 7:Unit 7.24. doi: 10.1002/0471142301.ns0724s60.
Abstract
DNA methylation is an epigenetic modification that is essential for the development and mature function of the central nervous system. Due to the relevance of this modification to the transcriptional control of gene expression, it is often necessary to examine changes in DNA methylation patterns with both gene and single-nucleotide resolution. Here, we describe an in-depth basic protocol for direct bisulfite sequencing of DNA isolated from brain tissue, which will permit direct assessment of methylation status at individual genes as well as individual cytosine molecules/nucleotides within a genomic region. This method yields analysis of DNA methylation patterns that is robust, accurate, and reproducible, thereby allowing insights into the role of alterations in DNA methylation in brain tissue.
摘要
DNA甲基化是一种表观遗传修饰，对中枢神经系统的发育和成熟功能至关重要。由于这种修饰与基因表达的转录控制相关，因此通常需要以基因和单核苷酸分辨率来检测DNA甲基化模式的变化。在这里，我们描述了一种用于对从脑组织中分离的DNA进行直接亚硫酸氢盐测序的深入基础方案，该方案将允许直接评估单个基因以及基因组区域内单个胞嘧啶分子/核苷酸的甲基化状态。这种方法能够对DNA甲基化模式进行稳健、准确且可重复的分析，从而有助于深入了解DNA甲基化改变在脑组织中的作用。
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