Methylation analysis by DNA immunoprecipitation (MeDIP).
Methylation analysis by DNA immunoprecipitation (MeDIP).
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DOI：
10.1007/978-1-60327-192-9_10
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发表时间：
2009
影响因子：
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通讯作者：
E. Vucic;Ian M. Wilson;Jennifer Campbell;W. Lam
中科院分区：
文献类型：
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作者：
E. Vucic;Ian M. Wilson;Jennifer Campbell;W. Lam
基因表达的表观遗传调控的改变在人类癌症和发育疾病中是常见的事件。CpG岛高甲基化和随之而来的基因沉默是观察到的，许多基因涉及多种功能和途径，在疾病状态下变得不受调控。因此，甲基组的比较图谱在疾病基因发现中是有用的。在全球范围内识别表观遗传学改变的能力对于理解与疾病进展平行的基因沉默模式至关重要。甲基化DNA免疫沉淀(MeDIP)是一种通过5‘-甲基胞嘧啶特异性抗体免疫沉淀来分离甲基化DNA片段的技术。然后，可以使用聚合酶链式反应方法以特定的位置分析甲基化DNA，或者通过与没有MeDIP浓缩的样品进行比较基因组杂交，以全基因组的方式进行分析。这篇文章描述了MeDIP和杂交到全基因组平铺路径BAC阵列的详细方案。
Alteration in epigenetic regulation of gene expression is a common event in human cancer and developmental disease. CpG island hypermethylation and consequent gene silencing is observed for many genes involved in a diverse range of functions and pathways that become deregulated in the disease state. Comparative profiling of the methylome is therefore useful in disease gene discovery. The ability to identify epigenetic alterations on a global scale is imperative to understanding the patterns of gene silencing that parallel disease progression.MethylatedDNAimmunoprecipitation (MeDIP) is a technique that isolates methylated DNA fragments by immunoprecipitating with 5′-methylcytosine-specific antibodies. The enriched methylated DNA can then be analyzed in a locus-specific manner using PCR assay or in a genome-wide fashion by comparative genomic hybridization against a sample without MeDIP enrichment. This article describes the detailed protocol for MeDIP and hybridization of MeDIP DNA to a whole-genome tiling path BAC array.