中国科学院机构知识库网格
Chinese Academy of Sciences Institutional Repositories Grid
Homogeneous and one-step fluorescent allele-specific PCR for SNP genotyping assays using conjugated polyelectrolytes

文献类型:期刊论文

作者Duan, Xinrui; Liu, Libing; Wang, Shu
刊名BIOSENSORS & BIOELECTRONICS
出版日期2009-03-15
卷号24期号:7页码:2095-2099
关键词Pcr Conjugated Polyelectrolytes Dna Snp Genotyping Fret Fluorescent Sensor
ISSN号0956-5663
DOI10.1016/j.bios.2008.10.027
英文摘要A new label-free, homogenous, sensitive and economical one-step method to detect SNP genotyping of genomic DNA has been developed by combining allele-specific PCR technique with water-soluble cationic conjugated polyelectrolytes (CCP). The amplification of target DNA and fluorescence detection steps are combined into one-step. The target DNA fragment containing a G allele site acts as PCR template. For the G allele-specific forward primer whose 3'-terminal base is complementary to the G allele template, after the first step of reverse primer extension, G allele-specific primer perfectly anneals with newly formed strand and the extension reaction of forward primer starts. During the extension, dGTP-F1 and dUTP-F1 are incorporated into extension chain in the presence of Taq DNA polymerase and more fluorescein-labeled PCR amplicons are yielded. Upon adding the CCP, strong electrostatic interactions between DNA and CCP bring them close and efficient FRET from CCP to fluorescein occurs. For the C allele-specific forward primer, less fluorescein-labeled PCR amplicons are yielded and inefficient FRET occurs. By triggering the change of emission intensity of CCP and fluorescein, it is possible to assay the SNP genotypes. In contrast to previous reports, this method does not require designing dye-labeled primers, and gel electrophoresis and isolation step after PCR were avoided in this homogenous method. The genotyping of 50 ng genomic DNA from human lung cancer cell is easily detected using our new method. These qualities will make the new detection system ideal for SNP genotyping. (C) 2008 Elsevier B.V. All rights reserved.
语种英语
WOS记录号WOS:000264791000046
出版者ELSEVIER ADVANCED TECHNOLOGY
源URL[http://ir.iccas.ac.cn/handle/121111/67563]  
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, Beijing Natl Lab Mol Sci, Key Lab Organ Solids, Beijing 100080, Peoples R China
推荐引用方式
GB/T 7714
Duan, Xinrui,Liu, Libing,Wang, Shu. Homogeneous and one-step fluorescent allele-specific PCR for SNP genotyping assays using conjugated polyelectrolytes[J]. BIOSENSORS & BIOELECTRONICS,2009,24(7):2095-2099.
APA Duan, Xinrui,Liu, Libing,&Wang, Shu.(2009).Homogeneous and one-step fluorescent allele-specific PCR for SNP genotyping assays using conjugated polyelectrolytes.BIOSENSORS & BIOELECTRONICS,24(7),2095-2099.
MLA Duan, Xinrui,et al."Homogeneous and one-step fluorescent allele-specific PCR for SNP genotyping assays using conjugated polyelectrolytes".BIOSENSORS & BIOELECTRONICS 24.7(2009):2095-2099.

入库方式: OAI收割

来源:化学研究所

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