Sørensen Anita L, Collas Philippe
Department of Biochemistry, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway.
Methods Mol Biol. 2009;567:249-62. doi: 10.1007/978-1-60327-414-2_16.
DNA methylation contributes to the regulation of long-term gene repression by enabling the recruitment of transcriptional repressor complexes to methylated cytosines. Several methods for detecting DNA methylation at the gene-specific and genome-wide levels have been developed. Methylated DNA immunoprecipitation, or MeDIP, consists of the selective immunoprecipitation of methylated DNA fragments using antibodies to 5-methylcytosine. The genomic site of interest can be detected by PCR, hybridization to DNA arrays, or by direct sequencing. This chapter describes the MeDIP protocol and quality control tests that should be performed throughout the procedure.
DNA甲基化通过促使转录抑制复合物募集到甲基化的胞嘧啶上，从而有助于长期基因抑制的调控。目前已开发出几种在基因特异性和全基因组水平检测DNA甲基化的方法。甲基化DNA免疫沉淀法（MeDIP），是利用针对5-甲基胞嘧啶的抗体对甲基化DNA片段进行选择性免疫沉淀。感兴趣的基因组位点可通过聚合酶链反应（PCR）、与DNA阵列杂交或直接测序来检测。本章介绍了MeDIP实验方案以及在整个过程中应进行的质量控制测试。
