Liu Chang-Gong, Spizzo Riccardo, Calin George Adrian, Croce Carlo Maria
Department of Molecular Virology, Immunology and Medical Genetics and Comprehensive Cancer Center, Ohio State University, Wiseman Hall Room 385, 400 12th Avenue, Columbus, OH 43210, United States.
Methods. 2008 Jan;44(1):22-30. doi: 10.1016/j.ymeth.2007.10.010.
After 12 years from its first application, microarray technology has become the reference technique to monitor gene expression of thousands of genes in the same experiment. In the past few years an increasing amount of evidence showed the importance of non-coding RNA (ncRNA) in different human diseases. The microRNAs (miRNAs) are one of the groups of ncRNA. They are small RNA fragments, 19-25 nucleotides long, with a main regulatory function on both protein coding genes and non-coding RNAs. The application of microarray platforms applied to miRNA profiling determined their deregulation in virtually all human diseases that have been studied. We previously developed a custom miRNA microarray platform, and here we describe the protocol we used to work with it including the oligo design strategy, the microarray printing protocol, the target-probe hybridization and the signal detection.
在首次应用12年后，微阵列技术已成为在同一实验中监测数千个基因表达的参考技术。在过去几年中，越来越多的证据表明非编码RNA（ncRNA）在不同人类疾病中的重要性。微小RNA（miRNA）是非编码RNA的一个类别。它们是长度为19 - 25个核苷酸的小RNA片段，对蛋白质编码基因和非编码RNA都具有主要的调节功能。应用于miRNA谱分析的微阵列平台的使用确定了它们在几乎所有已研究的人类疾病中的失调情况。我们之前开发了一个定制的miRNA微阵列平台，在此我们描述我们使用该平台的实验方案，包括寡核苷酸设计策略、微阵列打印方案、靶标 - 探针杂交以及信号检测。
