MicroRNA expression profiling using microarrays
MicroRNA expression profiling using microarrays
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DOI：
10.1038/nprot.2008.14
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发表时间：
2008-01-01
期刊：
影响因子：
14.8
通讯作者：
Croce, Carlo M.
中科院分区：
文献类型：
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作者：
Liu, Chang-Gong;Calin, George Adrian;Croce, Carlo M.
微阵列技术是一种强大的高通量工具，能够在一个实验中同时监测数十个平行处理的样品中数千个小的非编码RNA的表达。为了对正常和疾病样本（如癌症）的miRNA表达进行全基因组分析，并区分与诊断、预后和治疗干预相关的表达特征，我们在CodeLink平台上开发了一种独特的miRNA微阵列检测。miRNA阵列由一式两份打印的4，104个探针组成。该阵列可以同时分析来自474个人类和373个小鼠miRNA基因的1，500多个成熟miRNA及其相应的前体。本文详细介绍了本课题组开发的miRNA微阵列检测技术，包括miRNA寡核苷酸探针的设计、微阵列的制作和miRNA靶点的制备（通过逆转录总RNA）、微阵列上的靶点-探针杂交、信号检测和数据分析。该检测方法简单，易于标准化，并允许在24小时内对多达24个总RNA样品进行重复性分析。
Microarray technology is a powerful high-throughput tool capable of monitoring the expression of thousands of small noncoding RNAs at once within tens of samples processed in parallel in a single experiment. To conduct a genome-wide analysis of miRNA expression of normal and disease samples, such as cancer, and to distinguish expression signatures associated with diagnosis, prognosis and therapeutic interventions, we have developed a unique miRNA microarray assay on a CodeLink platform. The miRNA array consists of 4,104 probes printed in duplicate. This array can simultaneously profile more than 1,500 mature miRNAs and their corresponding precursors from 474 human and 373 mouse miRNA genes. The full protocol details of the miRNA microarray assay developed by our group are described here, including miRNA oligo probe design, array fabrication and miRNA target preparation (by reverse transcription of total RNA), target-probe hybridization on array, signal detection and data analysis. The assay is simple, can be easily standardized and allows the reproducible profiling of up to 24 total RNA samples within 24 h.