Optimization of oligonucleotide-based DNA microarrays -: art. no. e51
Optimization of oligonucleotide-based DNA microarrays -: art. no. e51
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DOI：
10.1093/nar/30.11.e51
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发表时间：
2002-06-01
影响因子：
14.9
通讯作者：
Valcárcel, J
中科院分区：
文献类型：
--
作者：
Relógio, A;Schwager, C;Valcárcel, J
基于寡核苷酸的DNA微阵列在基因表达和单核苷酸多态性分析中变得越来越有用。在这里，我们报告的灵敏度，特异性和动态范围的微阵列信号和它们的依赖标记和杂交条件，以及对寡核苷酸的长度，浓度，附着部分和纯度的系统研究。在这些实验中使用了一组受控的体外合成转录物和来自生物样品的RNA。提出了一种算法，允许有效地选择能够区分单核苷酸错配的寡核苷酸。各种应用程序的关键参数进行了讨论的基础上统计分析的结果。这些数据将有助于设计和标准化的定制微阵列适用于基因表达谱和测序分析。
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.