Fluorescent conjugated polymer-based FRET technique for detection of DNA methylation of cancer cells
文献类型：期刊论文
| 作者 | Feng, Fude; Liu, Libing; Wang, Shu |
| 刊名 | NATURE PROTOCOLS
 |
| 出版日期 | 2010 |
| 卷号 | 5期号:7页码:1255-1264 |
| ISSN号 | 1754-2189 |
| DOI | 10.1038/nprot.2010.79 |
| 英文摘要 | his protocol describes a homogeneous, convenient and sensitive DNA methylation detection method, using an optically amplifying cationic conjugated polymer (CCP, poly((1,4-phenylene)-2,7-[9,9-bis(6'-N,N,N-trimethyl ammonium)-hexyl fluorene] dibromide)). Genomic DNA from cancer cells is pretreated with a methylation-sensitive restriction endonuclease, followed by PCR amplification in the presence of fluorescein-labeled dNTP and Taq polymerase. The PCR only occurs for methylated DNA. DNA methylation of the gene sequence of interest is detected as a result of the fluorescence resonance energy transfer (FRET) between CCP and fluorescein that is incorporated into DNA. The methylated statuses of the p16, HPP1 and GALR2 promoters of five cancer cell lines (HT29, HepG2, A498, HL60 and M17) were assayed to provide an association study between the cancers and susceptibility genes, which shows great potential for early cancer diagnosis. This protocol simplifies previously available procedures by avoiding the need for primer labeling, isolation or purification steps, and sophisticated instruments. The assay takes about 20 h to obtain the methylated statuses of cancer cells. |
| 语种 | 英语 |
| WOS记录号 | WOS:000279404800003 |
| 出版者 | NATURE PUBLISHING GROUP |
| 源URL | [http://ir.iccas.ac.cn/handle/121111/69407]  |
| 专题 | 中国科学院化学研究所 |
| 通讯作者 | Wang, Shu |
| 作者单位 | Chinese Acad Sci, Inst Chem, Key Lab Organ Solids, Beijing Natl Lab Mol Sci, Beijing 100080, Peoples R China |
| 推荐引用方式 GB/T 7714 | Feng, Fude,Liu, Libing,Wang, Shu. Fluorescent conjugated polymer-based FRET technique for detection of DNA methylation of cancer cells[J]. NATURE PROTOCOLS,2010,5(7):1255-1264. |
| APA | Feng, Fude,Liu, Libing,&Wang, Shu.(2010).Fluorescent conjugated polymer-based FRET technique for detection of DNA methylation of cancer cells.NATURE PROTOCOLS,5(7),1255-1264. |
| MLA | Feng, Fude,et al."Fluorescent conjugated polymer-based FRET technique for detection of DNA methylation of cancer cells".NATURE PROTOCOLS 5.7(2010):1255-1264. |
入库方式： OAI收割
来源：化学研究所
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