Yang Yunfeng, Zhu Mengxia, Wu Liyou, Zhou Jizhong
Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA.
BMC Genomics. 2008 Sep 16;9 Suppl 2(Suppl 2):S5. doi: 10.1186/1471-2164-9-S2-S5.
Using genomic DNA as common reference in microarray experiments has recently been tested by different laboratories. Conflicting results have been reported with regard to the reliability of microarray results using this method. To explain it, we hypothesize that data processing is a critical element that impacts the data quality.
Microarray experiments were performed in a gamma-proteobacterium Shewanella oneidensis. Pair-wise comparison of three experimental conditions was obtained either with two labeled cDNA samples co-hybridized to the same array, or by employing Shewanella genomic DNA as a standard reference. Various data processing techniques were exploited to reduce the amount of inconsistency between both methods and the results were assessed. We discovered that data quality was significantly improved by imposing the constraint of minimal number of replicates, logarithmic transformation and random error analyses.
These findings demonstrate that data processing significantly influences data quality, which provides an explanation for the conflicting evaluation in the literature. This work could serve as a guideline for microarray data analysis using genomic DNA as a standard reference.
近期不同实验室已对在微阵列实验中使用基因组DNA作为通用参考进行了测试。关于使用该方法的微阵列结果的可靠性,已有相互矛盾的报道。为了解释这一现象,我们推测数据处理是影响数据质量的关键因素。
在γ-变形菌希瓦氏菌中进行了微阵列实验。通过将两个标记的cDNA样本共同杂交到同一阵列上,或者采用希瓦氏菌基因组DNA作为标准参考,对三种实验条件进行了成对比较。利用各种数据处理技术来减少两种方法之间的不一致性,并对结果进行了评估。我们发现,通过施加最少重复次数的约束、对数转换和随机误差分析,数据质量得到了显著提高。
这些发现表明,数据处理对数据质量有显著影响,这为文献中的矛盾评估提供了解释。这项工作可为以基因组DNA作为标准参考的微阵列数据分析提供指导。