| 荧光定量PCR检测细胞代谢综合征相关基因表达的优化 |
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| 引用本文： | 陈瑾歆,方定志.荧光定量PCR检测细胞代谢综合征相关基因表达的优化[J].中国热带医学,2010,10(9):1054-1056. |
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| 作者姓名： | 陈瑾歆 方定志 |
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| 作者单位： | 1. 川北医学院生物化学教研室,四川,南充,637000
2. 四川大学华西基础与法医学院,生物化学与分子生物教研室,四川,成都,610041 |
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| 摘 要： | 目的优化MRSG mRNA表达的Taqman荧光定量PCR检测反应体系并进行系统评价。方法设计荧光PCR适用的引物和探针,从模板、引物、探针、镁离子浓度等方面优化荧光定量PCR检测反应体系,评价优化系统的特异性和稳定性。结果确定系统的模板取样量为2μl,引物浓度为0.4μmol/L,探针浓度为0.2μmol/L,Mg~（2＋）浓度为4mM,建立了稳定的MRSG mRNA表达的Taqman荧光PCR检测方法。结论优化后的MRSG mRNA荧光PCR检测方法特异性和稳定性较好。
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| 关 键 词： | 代谢综合征相关基因 实时荧光定量PCR 系统优化 |
Optimization of detection of a novel metabolic syndrome related gene(MSRG)by taqman real-time PCR |
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| CHEN Jin-xin,FANG Ding-zhi.Optimization of detection of a novel metabolic syndrome related gene(MSRG)by taqman real-time PCR[J].China Tropical Medicine,2010,10(9):1054-1056. |
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| Authors: | CHEN Jin-xin FANG Ding-zhi |
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| Institution: | 1.Department of Biochemistry,North Sichuan Medical College,Nanchong 637000.,Sichuan,P.R.China) |
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| Abstract: | Aim To optimize the taqman real-time PCR reaction system for quantitative detection of the expression of MSRG mRNA and evaluate the results.Methods Specific primers and probes were designed and real-time PCR was used for d etection of cDNA sequence.Taqman real-time PCR reaction system from template,primer,probe and Mg~（2＋） concentration were optimized.Results Quantity of template was 2μl.The concentratons of primers,probes and Mg~（2＋） were 0.4 μmol/L,0.2 μmol/L and 4mM.Conclusion Via optimizing the real-time PCR assay a specific,reliable tool for detecting MRSG mRNA expression levels.was established. |
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| Keywords: | Metabolic syndrome related gene Taqman real-time PCR Systematic optimization |
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