ICCAS OpenIR
Fluorescent conjugated polymer-based FRET technique for detection of DNA methylation of cancer cells
Feng, Fude; Liu, Libing; Wang, Shu
2010
Source PublicationNATURE PROTOCOLS
ISSN1754-2189
Volume5Issue:7Pages:1255-1264
Abstracthis 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.
DOI10.1038/nprot.2010.79
Indexed BySCI
Language英语
WOS IDWOS:000279404800003
PublisherNATURE PUBLISHING GROUP
Citation statistics
Cited Times:54[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.iccas.ac.cn/handle/121111/69408
Collection中国科学院化学研究所
Corresponding AuthorWang, Shu
AffiliationChinese Acad Sci, Inst Chem, Key Lab Organ Solids, Beijing Natl Lab Mol Sci, Beijing 100080, Peoples R China
Recommended Citation
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.
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