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使用定量基因表达平台评估DNA微阵列结果。

Evaluation of DNA microarray results with quantitative gene expression platforms.

作者信息

Canales Roger D, Luo Yuling, Willey James C, Austermiller Bradley, Barbacioru Catalin C, Boysen Cecilie, Hunkapiller Kathryn, Jensen Roderick V, Knight Charles R, Lee Kathleen Y, Ma Yunqing, Maqsodi Botoul, Papallo Adam, Peters Elizabeth Herness, Poulter Karen, Ruppel Patricia L, Samaha Raymond R, Shi Leming, Yang Wen, Zhang Lu, Goodsaid Federico M

机构信息

Applied Biosystems, 850 Lincoln Centre Dr., Foster City, California 94404, USA.

出版信息

Nat Biotechnol. 2006 Sep;24(9):1115-22. doi: 10.1038/nbt1236.

Abstract

We have evaluated the performance characteristics of three quantitative gene expression technologies and correlated their expression measurements to those of five commercial microarray platforms, based on the MicroArray Quality Control (MAQC) data set. The limit of detection, assay range, precision, accuracy and fold-change correlations were assessed for 997 TaqMan Gene Expression Assays, 205 Standardized RT (Sta)RT-PCR assays and 244 QuantiGene assays. TaqMan is a registered trademark of Roche Molecular Systems, Inc. We observed high correlation between quantitative gene expression values and microarray platform results and found few discordant measurements among all platforms. The main cause of variability was differences in probe sequence and thus target location. A second source of variability was the limited and variable sensitivity of the different microarray platforms for detecting weakly expressed genes, which affected interplatform and intersite reproducibility of differentially expressed genes. From this analysis, we conclude that the MAQC microarray data set has been validated by alternative quantitative gene expression platforms thus supporting the use of microarray platforms for the quantitative characterization of gene expression.

摘要

基于微阵列质量控制(MAQC)数据集,我们评估了三种定量基因表达技术的性能特征,并将它们的表达测量结果与五个商业微阵列平台的结果进行了关联。对997种TaqMan基因表达分析、205种标准化逆转录(Sta)RT-PCR分析和244种QuantiGene分析的检测限、测定范围、精密度、准确度和倍数变化相关性进行了评估。TaqMan是罗氏分子系统公司的注册商标。我们观察到定量基因表达值与微阵列平台结果之间具有高度相关性,并且在所有平台中发现的不一致测量结果很少。变异性的主要原因是探针序列以及因此导致的靶标位置存在差异。变异性的第二个来源是不同微阵列平台检测弱表达基因的灵敏度有限且可变,这影响了差异表达基因的平台间和位点间再现性。通过该分析,我们得出结论,MAQC微阵列数据集已通过其他定量基因表达平台得到验证,从而支持使用微阵列平台对基因表达进行定量表征。

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