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通过微阵列进行甲基化分析。

Methylation analysis by microarray.

作者信息

Deatherage Daniel E, Potter Dustin, Yan Pearlly S, Huang Tim H-M, Lin Shili

机构信息

Human Cancer Genetics Program, The Ohio State University Comprehensive Cancer Center, The Ohio State University, Columbus, OH, USA.

出版信息

Methods Mol Biol. 2009;556:117-39. doi: 10.1007/978-1-60327-192-9_9.

DOI:10.1007/978-1-60327-192-9_9
PMID:19488875
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2838393/
Abstract

Differential methylation hybridization (DMH) is a high-throughput DNA methylation screening tool that utilizes methylation-sensitive restriction enzymes to profile methylated fragments by hybridizing them to a CpG island microarray. This array contains probes spanning all the 27,800 islands annotated in the UCSC Genome Browser. Herein we describe a DMH protocol with clearly identified quality control points. In this manner, samples that are unlikely to provide good read-outs for differential methylation profiles between the test and the control samples will be identified and repeated with appropriate modifications. The step-by-step laboratory DMH protocol is described. In addition, we provide descriptions regarding DMH data analysis, including image quantification, background correction, and statistical procedures for both exploratory analysis and more formal inferences. Issues regarding quality control are addressed as well.

摘要

差异甲基化杂交（DMH）是一种高通量DNA甲基化筛选工具，它利用甲基化敏感限制性内切酶，通过将甲基化片段与CpG岛微阵列杂交来分析甲基化片段。该阵列包含跨越UCSC基因组浏览器中注释的所有27,800个岛的探针。在此，我们描述了一种具有明确质量控制点的DMH方案。通过这种方式，将识别出不太可能为测试样本和对照样本之间的差异甲基化谱提供良好读数的样本，并进行适当修改后重复实验。文中描述了实验室DMH的详细步骤方案。此外，我们还提供了关于DMH数据分析的描述，包括图像定量、背景校正以及探索性分析和更正式推断的统计程序。同时也讨论了质量控制问题。

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