Construction of the standard curve for Real-time PCR detecting the expression of Angptl3 in rats

WANG Ji-hong; et al

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Construction of the standard curve for Real-time PCR detecting the expression of Angptl3 in rats

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- Construction of the standard curve for Real-time PCR detecting the expression of Angptl3 in rats
- Journal of Chongqing Medical University Vol. 34, Issue 5, Pages: 593-596(2009)
- 作者机构：
  
  重庆医科大学基础医学院生物化学与分子生物学教研室
- 作者简介：
- 基金信息：
- DOI：
  CLC： R450
- Published：2009，
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[1]王继红,黄家利,程梅,曾建涛,王欣.实时荧光定量PCR检测大鼠组织Angptl3表达的标准曲线构建[J].重庆医科大学学报,2009,34(05):593-596.

WANG Ji-hong, et al. Construction of the standard curve for Real-time PCR detecting the expression of Angptl3 in rats. [J]. Journal of Chongqing Medical University 34(5):593-596(2009)
[1]王继红,黄家利,程梅,曾建涛,王欣.实时荧光定量PCR检测大鼠组织Angptl3表达的标准曲线构建[J].重庆医科大学学报,2009,34(05):593-596. DOI：

WANG Ji-hong, et al. Construction of the standard curve for Real-time PCR detecting the expression of Angptl3 in rats. [J]. Journal of Chongqing Medical University 34(5):593-596(2009) DOI：

摘要

目的:采用实时荧光定量PCR（染料法）建立检测SD大鼠组织血管生成素样蛋白3（Angiopoietin-related protein3

Angptl3）表达量的标准曲线。方法:以SD大鼠肝组织总RNA为模板

通过RT-PCR获得目的DNA片段

用T-A克隆方法将其装载到质粒pUCm-T上

经过转化、质粒提取获得标准品

并以此为模板梯度稀释进行Real-time PCR反应

建立该方法的标准曲线。结果:Angptl3标准曲线相关系数为0.9976

Abstract

Objective:To construct the standard curve for real-time PCR detecting the tissue expression level of Angptl3 in SD rat.Methods:The total RNA of the SD rat’s liver was the template.We got lots of the Angptl3 partial DNA sequence by RT-PCR.The products were cloned into the pUCm-T vector

Then we diluted the solution of plasmid and used real-time PCR(SYBR GreenⅠ)to construct the standard curve.Results:The correlation coefficient of the standard curve was 0.997 6.The reaction efficiency was 0.862 3.The correlation and the amplify efficiency were good.Conclusion:It is successful to construct the standard curve for real-time PCR detecting the tissue expression level of Angptl3 in SD rat.

关键词

荧光定量PCRAngptl3标准曲线SD大鼠

Keywords

Real-time PCRAngptl3Standard curveSD rat

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