双标准曲线相对定量PCR试验原理与方法  

Theory and Method of Double-standard Curves Method of Relative Quantification PCR

作　　者：徐丽华[1,2] 刘春雷[1,2] 常玉梅[1] 梁利群[1] 刘金亮[1,2] 高国强[1,2] 韩启霞[1,2] 

机构地区：[1]中国水产科学院黑龙江水产研究所,哈尔滨150070 [2]上海海洋大学水产与生命学院,上海201306

出　　处：《生物技术通报》2011年第1期70-75,共6页Biotechnology Bulletin

基　　金：国家重点基础研究发展计划"973"项目(2010CB126305);中央级公益性科研院所基本科研业务费专项资金项目(2009HSYZX-SJ-05)

摘　　要：实时荧光定量PCR(FQ-PCR)是一种准确有效的核酸定量分析技术,具有易操作、高通量、高敏感性、高特异性、高度自动化和低污染等优点,并随新定量PCR仪及新操作方法的发展而得到广泛应用,但是,定量PCR的高敏感性特点使得实验操作严格而繁琐。阐述了一种改进的相对定量方法——双标准曲线法的试验原理和特点,描述了定量PCR体系的优化方式,探讨了试验误差分析方法及试验操作技巧,并就试验数据的处理方法进行讨论。试验证明,双标准曲线法是一种经济、简单而准确的定量方法。Real-time fluorescent quantitative polymerase chain reaction（FQ-PCR）is a nucleic acid quantification technique,has the advantage of simplicity of operation,high flux,high sensitivity,reliable specificity,highly automated and less polluted,and has been used widely with the introduction of new quantitative PCR instrumentation and operating methods.Real-time PCR experiments call for preciseness and redundancy because of high sensitivity.In this view,we discussed the experiment principle and experiment features of double-standard curves method——a improved relative quantification PCR method,amplify how to optimize PCR reaction system,then described the analytic approach of experimental error and experimental operative skills,at last,the means of dealing with experiment data.The results showed that double-standard curves method is economical,simple and accurate.

关 键 词：双标准曲线法 相对定量PCR 荧光扩增曲线 试验误差分析 归一化值 

分 类 号：R440[医药卫生—诊断学]