Section of Molecular Medicine, Department of Orthopedic Surgery, Rush University Medical Center, Chicago, IL 60612, USA.
Methods. 2010 Nov;52(3):213-7. doi: 10.1016/j.ymeth.2010.03.004. Epub 2010 Mar 19.
The methylated-CpG island recovery assay (MIRA) exploits the intrinsic specificity and the high affinity of a methylated-CpG-binding protein complex (MBD2B and MBD3L1) to methylated CpG dinucleotides in genomic DNA. The MIRA approach works on double-stranded DNA and does not depend on the application of methylation-sensitive restriction enzymes. It can be performed on a few hundred nanograms of genomic DNA. Recently, the MIRA technique has been used to profile DNA methylation patterns at a resolution of 100 base pairs along the entire genome of normal human B-lymphocytes. The MIRA method is compatible with microarray and next generation DNA sequencing approaches. We describe the principles and details of this method applied for methylation profiling of genomes containing methylated CpG sequences.
甲基化-CpG 岛回收分析 (MIRA) 利用甲基化-CpG 结合蛋白复合物 (MBD2B 和 MBD3L1) 对基因组 DNA 中甲基化 CpG 二核苷酸的固有特异性和高亲和力。MIRA 方法适用于双链 DNA,不依赖于甲基化敏感的限制性内切酶的应用。它可以在几百纳克的基因组 DNA 上进行。最近,MIRA 技术已被用于在整个正常人 B 淋巴细胞基因组上以 100 个碱基对的分辨率对 DNA 甲基化模式进行分析。MIRA 方法与微阵列和下一代 DNA 测序方法兼容。我们描述了该方法的原理和细节,该方法用于对包含甲基化 CpG 序列的基因组进行甲基化分析。