Real-Time PCR Technique and Its Application in Quantification of Plant Nucleic Acid Molecules

同步PCR技术及其在植物核酸分子定量中的应用(英文)

Real-time PCR is a closed DNA amplification system that skillfully integrates biochemical, photoelectric and computer techniques. Fluorescence data acquired once per cycle provides rapid absolute quantification of initial template copy numbers as PCR products are generated. This technique significantly simplifies and accelerates the process of producing reproducible quantification of nucleic acid molecules. It not only is a sensitive, accurate and rapid quantitative method, but it also provides an easier way to calculate the absolute starting copy number of nucleic acid molecules to be tested. Together with molecular bio-techniques, like microarray, real-time PCR will play a very important role in many aspects of molecular life science such as functional gene analysis and disease molecular diagnostics. This review introduces the detailed principles and application of the real-time PCR technique, describes a recently developed system for exact quantification of AUX/IAA genes In Arabidopsis, and discusses the problems with the real-time PCR process.

同步PCR是一种集生化、光电和计算机技术于一体的封闭式DNA扩增系统 ,采用荧光染料将扩增与检测过程结合在一起 ,实现了在PCR过程中在线显示PCR反应 ,通过检测荧光强度来绝对定量起始模板的拷贝数。该技术大大简化和加速了核酸分子的定量过程 ,不仅快速、灵敏、准确、重复性好 ,而且很容易计算出待测样品中核酸分子的绝对起始拷贝数。同微阵列等分子生物技术一起 ,同步PCR技术将会在功能基因解析和病害分子诊断等方面发挥重要作用。本综述除了介绍同步PCR技术的原理和应用外 ,还介绍了定量拟南芥Aux IAA基因的转录水平的实验 ,并就同步PCR操作过程中的问题进行了讨论。