Coman Diana, Gruissem Wilhelm, Hennig Lars
Plant Biotechnology, Department of Biology, ETH Zurich, Zurich, Switzerland.
Methods Mol Biol. 2013;1067:35-49. doi: 10.1007/978-1-62703-607-8_3.
Microarray technology is at present a standardized workflow for genome-wide expression analysis. Whole-genome tiling microarrays have emerged as an important platform for flexible and comprehensive expression profiling. In this chapter we describe a detailed standardized workflow for experiments assessing the transcriptome of Arabidopsis using tiling arrays and provide useful hints for critical steps from experimental design to data analysis. Although the protocol is optimized for AGRONOMICS1 arrays, it can readily be adapted to other tiling arrays. AGRONOMICS1 is the first platform that enables strand-specific expression analysis of the Arabidopsis genome with a single array. Moreover, it includes all perfect match probes from the original ATH1 array, allowing readily integration with the large existing ATH1 knowledge base. This workflow is designed for the analysis of raw data for any number of samples and it does not pose any particular hardware requirements.
微阵列技术目前是全基因组表达分析的标准化流程。全基因组平铺微阵列已成为灵活且全面的表达谱分析的重要平台。在本章中,我们描述了使用平铺阵列评估拟南芥转录组实验的详细标准化流程,并为从实验设计到数据分析的关键步骤提供有用的提示。尽管该方案是针对AGRONOMICS1阵列进行优化的,但它可以很容易地适用于其他平铺阵列。AGRONOMICS1是第一个能够通过单个阵列对拟南芥基因组进行链特异性表达分析的平台。此外,它包含了原始ATH1阵列的所有完全匹配探针,便于与现有的庞大ATH1知识库进行整合。此流程设计用于分析任意数量样本的原始数据,并且不提出任何特殊的硬件要求。