Gene expression studies using microarrays: Principles, problems, and prospects
Gene expression studies using microarrays: Principles, problems, and prospects
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DOI：
10.1152/advan.00043.2002
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发表时间：
2002-12-01
影响因子：
2.1
通讯作者：
Murphy, D
中科院分区：
文献类型：
--
作者：
Murphy, D
许多哺乳动物的基因组已经被测序，重要的下一步是通过使用微阵列对健康和疾病中所有转录单位的表达模式进行编目。这些发现项目对于我们理解控制发育、生理和病理过程的基因网络至关重要。然而，尽管令人兴奋，芯片技术的全部承诺尚未实现，因为该概念的表面简单掩盖了相当多的问题。微阵列技术是非常新的;方法仍在发展，共同的标准尚未建立，许多问题与实验设计和可变性仍有待充分理解和克服。这篇综述将描述微阵列实验的时间过程-从样品中分离RNA，靶制备，与微阵列探针杂交，数据捕获和生物信息学分析。对于每个阶段，竞争技术的优点和缺点进行了比较，并确定和讨论了固有的误差来源。
A number of mammalian genomes having been sequenced, an important next step is to catalog the expression patterns of all transcription units in health and disease by use of microarrays. Such discovery programs are crucial to our understanding of the gene networks that control developmental, physiological, and pathological processes. However, despite the excitement, the full promise of microarray technology has yet to be realized, as the superficial simplicity of the concept belies considerable problems. Microarray technology is very new; methodologies are still evolving, common standards have yet to be established, and many problems with experimental design and variability have still to be fully understood and overcome. This review will describe the time course of a microarray experiment-RNA isolation from sample, target preparation, hybridization to the microarray probe, data capture, and bioinformatic analysis. For each stage, the advantages and disadvantages of competing techniques are compared, and inherent sources of error are identified and discussed.