用于原代肌肉干细胞DNA甲基化分析的亚硫酸氢盐测序法
Bisulfite Sequencing for DNA Methylation Analysis of Primary Muscle Stem Cells.
作者信息
Miyata Kohei, Naito Masashi, Miyata Tomoko, Mokuda Sho, Asahara Hiroshi
机构信息
Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA, USA.
Department of Systems Bio Medicine, Tokyo Medical and Dental University, Tokyo, Japan.
出版信息
Methods Mol Biol. 2017;1668:3-13. doi: 10.1007/978-1-4939-7283-8_1.
Abstract
In skeletal muscle, DNA methylation contributes to the suppression of gene expression in several biological processes and diseases. A protocol for the detection of methylated cytosine was thus established based on methylation-sensitive enzymes, immunoprecipitation, and bisulfite conversion. DNA methylation analysis, with bisulfite conversion and sequencing, enables the quantification of methylation at each single base position. Here, we describe a basic method of bisulfite sequencing that can be used to analyze local DNA methylation status to confirm genome-wide DNA methylation analysis or correlation of gene expression regulatory mechanisms.
摘要
在骨骼肌中，DNA甲基化在多个生物学过程和疾病中有助于基因表达的抑制。因此，基于甲基化敏感酶、免疫沉淀和亚硫酸氢盐转化建立了一种检测甲基化胞嘧啶的方案。通过亚硫酸氢盐转化和测序进行的DNA甲基化分析能够对每个单碱基位置的甲基化进行定量。在此，我们描述了一种亚硫酸氢盐测序的基本方法，可用于分析局部DNA甲基化状态，以确认全基因组DNA甲基化分析或基因表达调控机制的相关性。
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