Relógio Angela, Schwager Christian, Richter Alexandra, Ansorge Wilhelm, Valcárcel Juan
Gene Expression Programme and Functional Genomics Technology and Instrumentation Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.
Nucleic Acids Res. 2002 Jun 1;30(11):e51. doi: 10.1093/nar/30.11.e51.
Oligonucleotide-based DNA microarrays are becoming increasingly useful for the analysis of gene expression and single nucleotide polymorphisms. Here we report a systematic study of the sensitivity, specificity and dynamic range of microarray signals and their dependence on the labeling and hybridization conditions as well as on the length, concentration, attachment moiety and purity of the oligonucleotides. Both a controlled set of in vitro synthesized transcripts and RNAs from biological samples were used in these experiments. An algorithm is presented that allows the efficient selection of oligonucleotides able to discriminate a single nucleotide mismatch. Critical parameters for various applications are discussed based on statistical analysis of the results. These data will facilitate the design and standardization of custom-made microarrays applicable to gene expression profiling and sequencing analyses.
基于寡核苷酸的DNA微阵列在基因表达分析和单核苷酸多态性研究中越来越有用。在此，我们报告了一项关于微阵列信号的灵敏度、特异性和动态范围及其对标记和杂交条件以及寡核苷酸长度、浓度、连接部分和纯度的依赖性的系统研究。在这些实验中，使用了一组受控的体外合成转录本以及来自生物样品的RNA。提出了一种算法，可有效选择能够区分单核苷酸错配的寡核苷酸。基于结果的统计分析讨论了各种应用的关键参数。这些数据将有助于适用于基因表达谱分析和测序分析的定制微阵列的设计和标准化。
