旨在建立嗜虫书虱的实时荧光定量PCR分析方法,为嗜虫书虱基因的定量分析提供技术支持。利用RT-PCR方法,从嗜虫书虱体内克隆获得了β-actin基因cDNA片段(GenBank登录号:FJ041117),该基因片段长度为822 bp,编码273个氨基酸残基(从第3个碱...旨在建立嗜虫书虱的实时荧光定量PCR分析方法,为嗜虫书虱基因的定量分析提供技术支持。利用RT-PCR方法,从嗜虫书虱体内克隆获得了β-actin基因cDNA片段(GenBank登录号:FJ041117),该基因片段长度为822 bp,编码273个氨基酸残基(从第3个碱基开始编码)。根据此β-actin基因的序列设计引物,建立了基于SYBR Green I染料技术的实时荧光定量PCR方法。建立的嗜虫书虱β-actin实时荧光定量PCR法扩增效率高、检测范围广、检测周期短,为β-actin作为内参基因进行嗜虫书虱功能基因的定量分析奠定了基础。展开更多
Real-time fluorescent quantitative PCR （RQ-PCR） is a detection method by adding fluorescent dye or fluorescent probe into the PCR reaction system, using fluorescent signal accumulation to monitor amplification react...Real-time fluorescent quantitative PCR （RQ-PCR） is a detection method by adding fluorescent dye or fluorescent probe into the PCR reaction system, using fluorescent signal accumulation to monitor amplification reactions of PCR reaction process, and finally the unknown template can be quantitatively analyzed through the standard curve. So the detection level of PCR has improved from the qualitative to the quantitative. In order to provide a theoretical reference for further application, the principle, classification, advantages and disadvantages of RQ-PCR were intro- duced, and its application and progress in plants in recent years were reviewed.展开更多
文摘旨在建立嗜虫书虱的实时荧光定量PCR分析方法,为嗜虫书虱基因的定量分析提供技术支持。利用RT-PCR方法,从嗜虫书虱体内克隆获得了β-actin基因cDNA片段(GenBank登录号:FJ041117),该基因片段长度为822 bp,编码273个氨基酸残基(从第3个碱基开始编码)。根据此β-actin基因的序列设计引物,建立了基于SYBR Green I染料技术的实时荧光定量PCR方法。建立的嗜虫书虱β-actin实时荧光定量PCR法扩增效率高、检测范围广、检测周期短,为β-actin作为内参基因进行嗜虫书虱功能基因的定量分析奠定了基础。
基金Supported by National Natural Science Foundation of China(31260406)Natural Science Fund Project of Inner Mongolia(2012MS0502)~~
文摘Real-time fluorescent quantitative PCR （RQ-PCR） is a detection method by adding fluorescent dye or fluorescent probe into the PCR reaction system, using fluorescent signal accumulation to monitor amplification reactions of PCR reaction process, and finally the unknown template can be quantitatively analyzed through the standard curve. So the detection level of PCR has improved from the qualitative to the quantitative. In order to provide a theoretical reference for further application, the principle, classification, advantages and disadvantages of RQ-PCR were intro- duced, and its application and progress in plants in recent years were reviewed.
