Bustin S A, Benes V, Nolan T, Pfaffl M W
Institute of Cellular and Molecular Science, Barts and the London, Queen Mary's School of Medicine and Dentistry, University of London, London, UK.
J Mol Endocrinol. 2005 Jun;34(3):597-601. doi: 10.1677/jme.1.01755.
The real-time reverse transcription polymerase chain reaction (RT-PCR) uses fluorescent reporter molecules to monitor the production of amplification products during each cycle of the PCR reaction. This combines the nucleic acid amplification and detection steps into one homogeneous assay and obviates the need for gel electrophoresis to detect amplification products. Use of appropriate chemistries and data analysis eliminates the need for Southern blotting or DNA sequencing for amplicon identification. Its simplicity, specificity and sensitivity, together with its potential for high throughput and the ongoing introduction of new chemistries, more reliable instrumentation and improved protocols, has made real-time RT-PCR the benchmark technology for the detection and/or comparison of RNA levels.
实时逆转录聚合酶链反应（RT-PCR）使用荧光报告分子来监测PCR反应每个循环中扩增产物的生成。这将核酸扩增和检测步骤整合到一个均相测定中，无需进行凝胶电泳来检测扩增产物。使用合适的化学方法和数据分析，无需进行Southern印迹或DNA测序来鉴定扩增子。其简单性、特异性和敏感性，以及高通量的潜力，加上不断引入新的化学方法、更可靠的仪器和改进的方案，使实时RT-PCR成为检测和/或比较RNA水平的基准技术。
