| 实时定量PCR检测内质网应激相关基因方法的建立 |
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| 引用本文： | 王夏,沈玉君,徐元宏,沈玉先.实时定量PCR检测内质网应激相关基因方法的建立[J].安徽医科大学学报,2012,47(8):988-991. |
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| 作者姓名： | 王夏 沈玉君 徐元宏 沈玉先 |
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| 作者单位： | 安徽医科大学药学院,合肥,230032;安徽医科大学基础医学院,合肥,230032;安徽医科大学教育部重要遗传病基因资源利用重点实验室,省部共建,合肥,230032;安徽医科大学第一附属医院检验科,合肥,230022;安徽医科大学基础医学院,合肥,230032;安徽医科大学教育部重要遗传病基因资源利用重点实验室,省部共建,合肥,230032 |
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| 摘 要： | 从人外周血白细胞中扩增目的基因片段,构建标准品质粒,应用SYBR GreenⅠ荧光染料进行实时定量PCR,检测内质网应激相关基因的拷贝数。所建立的实时定量PCR方法在起始模板量106～1013拷贝/μl范围内,荧光信号达到阈值的循环数Ct值与模板浓度线性关系良好(R2=0.991 9),是检测内质网应激相关基因有效的手段。
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| 关 键 词： | 实时定量PCR SYBR GreenⅠ 内质网应激 |
Quantitative PCR detection of endoplasmic reticulum stress associated genes |
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| Institution: | Wang Xia1,2,Shen Yujun3,Xu Yuanhong4,et al(1School of Pharmacy,2School of Basic Medicine,Anhui Medical University,3Key Laboratory of Gene Resource Utilization for Genetic Diseases of Educational Ministry and Anhui Province,Hefei 230032; 4Dept of Clinical Laboratory,The First Affiliated Hospital of Anhui Medical University,Hefei 230022) |
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| Abstract: | Target gene fragments were amplified from human peripheral white blood cells,and the standard plasmids were constructed to detect ER stress-associated genes using real time quantitative polymerase chain reaction(PCR) in the present of SYBR GreenⅠ.There was a significantly linear correlation(R2=0.991 9) between the threshold Ct and the template DNA concentration ranging from 106 to 1013 copies.It is an effective method to detect the mRNA expression level of ER stress-associated genes. |
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| Keywords: | real-time quantitative PCR SYBR GreenⅠ ER stress |
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