实时荧光定量PCR检测犬GAPDH基因  

GAPDH Gene measured by real-time fluorescent quantitative PCR

作　　者：李佳[1,2] 姜代勋[1] 陈益山[1,2] 任超[1,2] 陈丽[1,2] 张岩[1,2] 陈武[1,2] 

机构地区：[1]北京农学院兽医学(中医药)北京市重点实验室,北京102206 [2]北京农学院动物科学技术系,北京102206

出　　处：《北京农学院学报》2010年第1期33-36,共4页Journal of Beijing University of Agriculture

基　　金：国家自然科学基金项目(30671542)

摘　　要：建立犬3-磷酸甘油醛脱氢酶基因的qPCR检测体系,为犬功能基因mRNA水平的定量分析提供有用的方法学基础。对反应体系进行优化,建立了基于SYBR GreenI染料技术的real-time PCR方法。结果表明,其标准曲线相关系数达到0.999,扩增效率为105.8%,熔解曲线为特异性单峰,组内变异系数（CV%）为0.25-2.8,组间变异系数（CV%）为1.31-2.85。该研究所建立的荧光定量RT-PCR法灵敏、稳定、高效,为犬GAPDH基因作为内参基因用于犬相关基因定量表达分析奠定了基础。This study was designed to establish real-time PCR detection system of canine glyceraldehydes-3-phosphate dehydrogenase（GAPDH） gene,providing the useful methodology basis for quantitative analysis in the level of mRNA of canine functional genes.After optimization of the reaction system,the real-time PCR method based on SYBR Green I dye-based technology was established.The following results were obtained：the good correlation coefficient of 0.999,the amplification efficiency of qPCRs at 105.8%.The melting curve presented a single peak.The intra-assay variability（CV%） were 0.25~2.8,and the inter-assay variability（CV%）were 1.31~2.85.The results in this study indicated that the qPCR had advantages of high sensitivity,repeatability and reproducibility.The assay provide the basis for GAPDH gene of canine as a reference gene in quantitative analysis of mRNA expression.

关 键 词：犬 GAPDH基因 qPCR 

分 类 号：S818.9[农业科学—畜牧学]