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题名: Methylation analysis of hMLH1 gene promoter by a bisulfate-sensitive single-strand conformation polymorphism-capillary electrophoresis method
作者: Shi, Xianzhe;  Li, Jianhua;  Zhao, Chunxia;  Lv, Shen;  Xu, Guowang
关键词: DNA methylation ;  hMLH1 promoter ;  bisulfite-single strand conformation polymorphism ;  capillary electrophoresis
刊名: BIOMEDICAL CHROMATOGRAPHY
发表日期: 2006-08-01
DOI: 10.1002/bmc.606
卷: 20, 期:8, 页:815-820
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Biochemistry & Molecular Biology ;  Chemistry, Analytical ;  Pharmacology & Pharmacy
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry ;  Pharmacology & Pharmacy
英文摘要: DNA methylation is an important epigenetic modification that alters transcription in those genes containing CpG islands. In this report, a novel DNA methylation analysis method was developed employing bisulfite-single strand conformation polymorphism combined with capillary electrophoresis (bisulfite-SSCP-CE). During the bisulfite treatment of genomic DNA, a high concentration of sodium bisulfite (4.8 mol/L) was preferred in order to shorten reaction time and minimize template degradation. The methylated and unmethylated ssDNA of hMLH1 promoter were simultaneously separated under the optimized CE conditions, including 6% SLPA with 10% glycerol as sieving medium, 25 degrees C as separation temperature and 12 kV as running voltage. The heterogeneous methylation of hMLH1 promoter was identified in 13 of 64 colorectal cancer patients. Moreover, hMLH1 promoter methylation had a significant relationship with protein expression loss and increased with the age of patients. Our results indicated that DNA methylation analysis for a large number of clinical samples would be facilitated by use of the bisulfite-SSCP-CE method. Copyright (c) 2005 John Wiley & Sons, Ltd.
关键词[WOS]: MUTATION DETECTION ;  CPG ;  EXPRESSION ;  ASSAY ;  PCR
语种: 英语
WOS记录号: WOS:000239541300020
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/140042
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Natl Chromatog R&A Ctr, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
2.Dalian Med Univ, Secondary Affiliated Hosp, Dalian 116027, Peoples R China

Recommended Citation:
Shi, Xianzhe,Li, Jianhua,Zhao, Chunxia,et al. Methylation analysis of hMLH1 gene promoter by a bisulfate-sensitive single-strand conformation polymorphism-capillary electrophoresis method[J]. BIOMEDICAL CHROMATOGRAPHY,2006,20(8):815-820.
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