荧光定量PCR检测细胞代谢综合征相关基因表达的优化

Optimization of detection of a novel metabolic syndrome related gene (MSRG) by taqman real-time PCR

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。

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.