Peano Clelia, Severgnini Marco, Cifola Ingrid, De Bellis Gianluca, Battaglia Cristina
Institute for Biomedical Technologies, National Research Council, Milan, Italy.
Expert Rev Mol Diagn. 2006 May;6(3):465-80. doi: 10.1586/14737159.6.3.465.
The increasing use of microarray expression profiling to study the molecular biology of cancer and the cellular physiology of difficult-to-isolate cell types has led to a need for methods that accurately and precisely amplify small quantities of RNA. The purpose of this review is to provide an overview of the existing methods for transcriptome amplification and to define the parameters for comparing different amplification methods. The authors propose a standardized protocol for the assessment and evaluation of amplification methods, focusing on a new whole-transcriptome amplification kit, which amplifies total RNA into cDNA fragments. Reproducibility and reliability of the method were analyzed and discussed using both quantitative real-time PCR and a high-density oligonucleotide microarray platform.
微阵列表达谱分析在研究癌症分子生物学和难以分离的细胞类型的细胞生理学方面的应用日益广泛，这就需要能够准确、精确地扩增少量RNA的方法。本综述的目的是概述现有的转录组扩增方法，并确定比较不同扩增方法的参数。作者提出了一种评估和评价扩增方法的标准化方案，重点介绍了一种新的全转录组扩增试剂盒，该试剂盒可将总RNA扩增成cDNA片段。使用定量实时PCR和高密度寡核苷酸微阵列平台对该方法的重现性和可靠性进行了分析和讨论。
