摘要
本研究旨在建立定量检测小鼠-βactin mRNA表达水平的实时荧光定量PCR方法,为小鼠相关基因表达水平的分析提供方法。根据GenBank中登录的小鼠-βactin mRNA序列,针对保守区域设计并合成1对引物,建立了基于SYBR GreenⅠ技术的小鼠-βactin mRNA荧光定量PCR方法。结果表明,该方法扩增效率高（97.7%）,定量线性范围广（9.3×102~9.3×107拷贝/反应）,可重复性强,无非特异性扩增和引物二聚体,最小检出量为9.3拷贝/反应。试验结果为-βactin作为内参基因用于小鼠相关基因转录水平分析提供了基础。
The aim of present study is to establish a real-time PCR for quantitative analysis of mouse β-actin mRNA expression.A set of primer was designed according to the mouse β-actin mRNA sequence in GenBank.And a SYBR Green Ⅰ-based reverse real-time PCR（RRT-PCR） was successfully developed.The results showed that RRT-PCR assay established had a high amplification efficiency（97.7%）,wide linear range（9.3×102 to 9.3×107 copies/reaction）,good reproducibility and specificity.The detection limit was 9.3 copies/reaction.The results provided a basis for use of β-actin as a reference gene in gene expression analysis in mouse.
出处
《中国畜牧兽医》
CAS
北大核心
2011年第7期127-130,共4页
China Animal Husbandry & Veterinary Medicine
基金
四川省科技条件平台建设项目
