Quantitative assessment of DNA microarrays - Comparison with Northern blot analyses
Quantitative assessment of DNA microarrays - Comparison with Northern blot analyses
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DOI：
10.1006/geno.2000.6427
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发表时间：
2001-01-01
期刊：
影响因子：
4.4
通讯作者：
Yamanaka, S
中科院分区：
文献类型：
--
作者：
Taniguchi, M;Miura, K;Yamanaka, S
DNA微阵列是一种强大的技术，可以提供数千个基因的表达谱。然而，较少注意到其数量方面。在这项研究中，我们构建了一个小规模的DNA微阵列，包含84个基因，并表征其定量方面。该微阵列分析显示，在小鼠胚胎干细胞分化过程中，17个基因被诱导，而8个基因被抑制至少两倍。当用相同的荧光染料组合重复进行探针标记时，微阵列产生一致的数据（相关系数= 0.991）。相反，当用染料的反向组合重复时，数据不太一致（相关系数= 0.945）。染料组合的影响在几个基因中特别明显。总RNA（15 μ g）和poly（A）RNA（0.5 μ g）显示出相当的灵敏度，并产生基本相同的数据（相关系数= 0.983）。DNA微阵列的灵敏度略低于北方印迹分析。在大多数基因中，两种方法获得的数据是一致的。然而，在46个基因中，有4个基因的表达变化，DNA芯片未能检测到所揭示的北方印迹。这些数据表明，DNA微阵列提供的定量数据与一般的北方印迹相当，但在分析数据时必须考虑一些问题。(C)北京：科学出版社.
DNA microarray is a powerful technology that provides the expression profile of thousands of genes. However, less attention has been paid to its quantitative aspect. In this study, we constructed a small-scale DNA microarray that contains 84 genes and characterized its quantitative aspect. Analyses with this microarray showed that 17 genes were induced, whereas 8 genes were suppressed at least twofold during the differentiation of mouse embryonic stem cells. When repeated with the same combination of fluorescent dyes for probe labeling, the microarray produced consistent data (correlation coefficient = 0.991). In contrast, data were less consistent when repeated with the reverse combination of dyes (correlation coefficient = 0.945). The effect of dye combination was particularly evident in several genes. Total RNA (15 mug) and poly(A) RNA (0.5 mug) showed comparable sensitivity and produced essentially identical data (correlation coefficient = 0.983). The sensitivity of the DNA microarrays was slightly inferior to that of Northern blot analyses. In most genes, data obtained with the two methods were consistent. However, in 4 of 46 genes compared, DNA microarrays failed to detect the expression changes that were revealed by Northern blot. These data demonstrated that DNA microarrays provide quantitative data comparable to Northern blot in general, but a few issues must be considered when analyzing data. (C) 2001 Academic Press.