ICON probes: synthesis and DNA methylation typing.
ICON probes: synthesis and DNA methylation typing.
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ICON 探针：合成和 DNA 甲基化分型。
DOI：
10.1002/0471142700.nc0807s47
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Okamoto A.
中科院分区：
文献类型：
--
作者：
Tainaka K;Okamoto A.
DNA甲基化和去甲基化分别显著影响基因表达的失活和激活过程。测定DNA甲基化的位置和频率对于阐明细胞分化和癌变的机制具有重要意义，并可能成为一种有用和有效的癌症诊断指标。我们已经开发了一种人工DNA探针，其诱导长DNA序列中靶胞嘧啶的甲基化检测反应（ICON探针）。该人工DNA允许快速检测连接在靶胞嘧啶的C5位置处的甲基。此外，在该反应中不存在基因组DNA的非特异性切割。ICON探针还有助于使用少量基因组DNA样品定量靶胞嘧啶处的甲基化。本单元提供了合成联吡啶修饰的腺苷亚磷酰胺和制备ICON探针的程序。此外，还提供了利用ICON探针通过定量PCR进行甲基化定量实验的方案。方案核酸化学47：8.7.1 - 8.7.17。© 2011年由John Wiley & Sons，Inc.
DNA methylation and demethylation significantly affect the deactivation and activation processes of gene expression, respectively. The determination of the location and frequency of DNA methylation is important for the elucidation of the mechanisms of cell differentiation and carcinogenesis and may be a useful and effective index for cancer diagnosis. We have developed an artificial DNA probe that induces a methylation detection reaction of a target cytosine in a long DNA sequence (ICON probe). This artificial DNA allows the rapid detection of a methyl group attached at the C5 position of the target cytosine. In addition, there is no nonspecific cleavage of genomic DNA in this reaction. The ICON probe also facilitates the quantification of methylation at the target cytosine using a small amount of genomic DNA sample. This unit provides a procedure for synthesizing bipyridine‐modified adenosine phosphoramidite and preparation of ICON probes. Additionally, the protocol for the methylation quantification experiments by quantitative PCR utilizing ICON probes is also presented.Curr. Protoc. Nucleic Acid Chem. 47:8.7.1‐8.7.17. © 2011 by John Wiley & Sons, Inc.