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Updated: Jun 14, 2026

DNA Methylation: Bisulphite Modification and Analysis

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

全基因组双硫酸盐测序协议用于分析全基因组DNA甲基化和基甲基化模式的单核酸分辨率

David Derbala¹, Abel Garnier¹, Eric Bonnet¹

¹Centre National de Recherche en Génomique Humaine, CEA - Institut de Biologie François Jacob, Université Paris -Saclay, Evry, France.

Methods in molecular biology (Clifton, N.J.)

|July 16, 2024

概括

这项研究提出了一种全基因组二硫酸盐测序 (WGBS) 的新协议,用于分析DNA甲基化和基甲基化. 该方法区分这些表观遗传标记,为个性化医学研究提供了强大的工具.

科学领域:

表观遗传学在表观遗传学中,表观遗传学是指表观遗传学.
基因组学就是基因组学.
分子生物学分子生物学

背景情况:

基因甲基化和基甲基化是研究生物和临床应用的关键表观遗传修饰.
全基因组二硫酸盐测序 (WGBS) 是DNA甲基化分析的黄金标准,但无法区分基甲基化.
区分5-甲基细胞因和5-基甲基细胞因需要额外的步骤.

研究的目的:

为准备测序库提供详细的协议,用于全面的全基因组DNA甲基化和/或基甲基化分析.
为了在单个工作流程中量化区分5-甲基细胞因和5-基甲基细胞因水平.
为有限的DNA输入提供适用于有限的用户友好的方法.

主要方法:

一个使用商业套件测序库准备从碎片化DNA的协议.
在二硫酸盐转换之前,甲基化适配器的结合.
包括氧化双硫酸盐测序 (OxBs-Seq) 的氧化步骤,以区分甲基化和基甲基化.

主要成果:

成功生成了两种类型的测序库:一个用于总修饰的细胞因子,一个仅用于甲基化细胞因子.
通过比较两种类型的库的结果,可以准确地推断出基甲基化水平.
该协议在人类和植物样本中产生了可靠和可重复的结果.

结论:

描述的协议允许使用WGBS对DNA甲基化和基甲基化进行全面的定量分析.
这种方法通过详细的表观基因组分析,为推进个性化医疗提供了宝贵的工具.
该协议是高效的,可复制的,适用于不同的物种.

关键词:

双硫酸盐的转化转化通过DNA甲基化.氧甲基化氧甲基化.低输入的输入量很低.在OxBS中使用OxBS.氧化双硫酸盐序列的排序真甲基 (TrueMethyl) 是一种真甲基.超低的甲基-seqq.这是WGBS的WGBS.全基因组双硫酸盐测序

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Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

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