| 无标记探针技术检测IL-6基因启动子单核苷酸多态性 |
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| 引用本文： | 郜莉娜,尤崇革,潘云燕,李光迪,王芳芳.无标记探针技术检测IL-6基因启动子单核苷酸多态性[J].第四军医大学学报,2009(16):1484-1487. |
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| 作者姓名： | 郜莉娜 尤崇革 潘云燕 李光迪 王芳芳 |
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| 作者单位： | 兰州大学第二医院检验科; |
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| 摘 要： | 目的：探讨无标记探针技术检测IL-6基因启动子单核苷酸多态性（SNP）的实用性．方法：采用新型荧光染料EvaGreen和无标记探针技术对IL-6基因启动子SNP进行基因分型．以其-597G〉A位点为例设计PCR扩增引物和无标记探针，按PCR扩增效率和产物特异性进行Mg^2＋浓度、不对称PCR引物比例以及模板浓度等条件的优化．并用此优化体系基因分型100例临床标本，杂合型与突变型以测序验证．结果：结果显示EvaGreen荧光PCR检测体系是最适Mg^2＋浓度为2．5mmol／L，不对称PCR引物比例为0．1／0．5umol／L，模板浓度为10ng／10uL的两步法不对称PCR．该体系对100例样本进行IL-6基因～597G〉A位点基因分型，发现7例杂合型和1例突变型，经测序与检测结果一致．结论：无标记探针技术检测基因SNP是一种值得推广的简便易行、低成本、常规化的基因分型方法．
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| 关 键 词： | 白细胞介素6 EvaGreen 无标记探针 多态性 单核苷酸 |
Unlabeled probe genotyping for detection of single nucleotide polymorphism of IL-6 promoter |
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| GAO Li-Na,YOU Chong-Ge,PAN Yun-Yan,LI Guang-Di,WANG Fang-Fang Clinical Laboratory,Second Hospital of Lanzhou University,Lanzhou ,China.Unlabeled probe genotyping for detection of single nucleotide polymorphism of IL-6 promoter[J].Journal of the Fourth Military Medical University,2009(16):1484-1487. |
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| Authors: | GAO Li-Na YOU Chong-Ge PAN Yun-Yan LI Guang-Di WANG Fang-Fang Clinical Laboratory Second Hospital of Lanzhou University Lanzhou China |
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| Institution: | GAO Li-Na,YOU Chong-Ge,PAN Yun-Yan,LI Guang-Di,WANG Fang-Fang Clinical Laboratory,Second Hospital of Lanzhou University,Lanzhou 730030,China |
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| Abstract: | AIM:To study the practicality of using unlabeled probe genotyping for detecting single nucleotide polymorphism(SNP)of IL-6 gene promoter.METHODS:SNP of IL-6 gene promoter was genotyped with a new fluorescent dye EvaGreen and the latest unlabeled probe technique,and the PCR primers and unlabeled probe were designed on the SNP site-597G>A of IL-6 gene promoter.The PCR amplification system was optimized,including the concentration of Mg2+,the ratio of asymmetric PCR primer,the lowest concentration of template ... |
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| Keywords: | intekleukin-6 EvaGreen unlabeled probes polymorphism single nucleotide |
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