DNA methylation profiling using the methylated-CpG island recovery assay (MIRA).

DNA methylation profiling using the methylated-CpG island recovery assay (MIRA).
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DOI:
10.1016/j.ymeth.2010.03.004
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发表时间:
2010-11
期刊:
影响因子:
4.8
通讯作者:
Pfeifer, Gerd P.
Pfeifer, Gerd P.
中科院分区:
生物学3区
文献类型:
--
作者:
Rauch, Tibor A.;Pfeifer, Gerd P.

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甲基化CpG岛恢复试验(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性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技术已被用于沿着正常人B淋巴细胞的整个基因组以100个碱基对的分辨率描绘DNA甲基化模式。MIRA方法与微阵列和下一代DNA测序方法兼容。我们描述了这种方法的原理和细节,适用于含有甲基化CpG序列的基因组的甲基化分析。
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.
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