动物医学进展Issue(12)：64-68,5.
小鼠 Bax 基因实时荧光定量 PCR 标准品质粒的构建
Establishment of Standard Curve for mouse Bax Gene Recombinant Plasmid by Real-time qPCR
摘要
Abstract
The aim of the present study was to construct the standard plasmid and standard curve of real-time PCR for the quantitation of mRNA expression level of the mouse Bax gene.A pairs of specific primer were designed based on the conserved coding region from GeneBank.Total RNA was extracted from N2A cell and the fragments of target gene about 162bp were amplified by RT-PCR.Plasmid DNA was purified and identified by PCR amplification and sequencing after recycling,purification and transformation.The positive plasmids as standard template were entered into the SYBR Green Ⅰ real-time qPCR steps after 10 times dilution.The standard curve and regressive curve were generated automatically.The results showed that no specific amplicons was found according to the derived melting curve of the real-time PCR products, indicating the specification of the present primers.The correlation coefficients of the standard were 0.999, suggesting the strong linear relationship between the groups.The construction of the standard plasmid and standard curve were established for Bax gene.关键词
Bax基因/实时荧光定量PCR/标准品质粒/标准曲线Key words
Bax gene/real-time qPCR/standard plasmid/standard curve分类
生物科学引用本文复制引用
潘博,蔡青,鲁强,付文卓,王继宏,潘耀谦..小鼠 Bax 基因实时荧光定量 PCR 标准品质粒的构建[J].动物医学进展,2013,(12):64-68,5.基金项目
河南省重点科技攻关项目 （）
