Peterson Samuel M, Freeman Jennifer L
School of Health Sciences, Purdue University.
J Vis Exp. 2009 Aug 10(30):1471. doi: 10.3791/1471.
Gene microarray technology permits quantitative measurement and gene expression profiling of transcript levels on a genome-wide basis. Gene microarray technology is used in numerous biological disciplines in a variety of applications including global gene expression analysis in relation to developmental stage, to a disease state, and in toxic responses. Herein, we include a demonstration of global gene expression analysis using a comprehensive zebrafish-specific oligonucleotide microarray platform. The zebrafish expression microarray platform contains 385,000 probes, 60 base pairs in length, interrogating 37,157 targets with up to 12 probes per target. For this platform, all cDNA and genomic information available for the zebrafish was collected from various genomic databases including Ensembl (http://www.ensembl.org), VEGA (http://vega.sanger.ac.uk), UCSC (http://genome.ucsc.edu), and ZFIN (http://www.zfin.org). As a result this expression array provides complete coverage of the current zebrafish transcriptome. The zebrafish expression microarray was printed by Roche NimbleGen (Madison, WI). This technical demonstration includes the fluorescent labeling of a cDNA product, hybridization of the labeled cDNA product to the microarray platform, and array scanning for signal acquisition using the one color analysis strategy.
基因微阵列技术可在全基因组范围内对转录水平进行定量测量和基因表达谱分析。基因微阵列技术在众多生物学学科中有着广泛应用，包括与发育阶段、疾病状态以及毒性反应相关的全基因组基因表达分析。在此，我们展示了使用全面的斑马鱼特异性寡核苷酸微阵列平台进行全基因组基因表达分析的过程。斑马鱼表达微阵列平台包含38.5万个探针，每个探针长度为60个碱基对，可检测37157个靶点，每个靶点最多有12个探针。对于该平台，所有可获取的斑马鱼cDNA和基因组信息均来自包括Ensembl（http://www.ensembl.org）、VEGA（http://vega.sanger.ac.uk）、UCSC（http://genome.ucsc.edu）和ZFIN（http://www.zfin.org）在内的各种基因组数据库。因此，该表达阵列可完全覆盖当前的斑马鱼转录组。斑马鱼表达微阵列由罗氏NimbleGen公司（威斯康星州麦迪逊市）印制。本技术演示包括cDNA产物的荧光标记、标记后的cDNA产物与微阵列平台的杂交以及使用单色分析策略进行阵列扫描以获取信号。
