A genome-wide scalable SNP genotyping assay using microarray technology
A genome-wide scalable SNP genotyping assay using microarray technology
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DOI：
10.1038/ng1547
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发表时间：
2005-05-01
期刊：
影响因子：
30.8
通讯作者：
Chee, MS
中科院分区：
文献类型：
--
作者：
Gunderson, KL;Steemers, FJ;Chee, MS
寡核苷酸探针阵列能够对单个cDNA样本的基因表达水平进行大规模平行分析。将微阵列技术应用于基因组DNA分析一直受到整个人类基因组序列复杂性的阻碍。一种稳健的、单碱基分辨率的直接基因组检测方法将扩大微阵列技术的应用范围。我们开发了一种基于阵列的全基因组基因分型检测方法，该方法不需要聚合酶链反应（PCR），并且能够实现几乎无限的多重检测。该检测方法通过将皮摩尔浓度的全基因组扩增DNA与阵列探针的特异性杂交，以及等位基因特异性引物延伸和信号放大相结合，实现了高信噪比。作为原理验证，我们对数百个先前已确定特征的单核苷酸多态性（SNP）进行了基因分型。其转化率、识别率和准确性与高性能的基于PCR的基因分型检测方法相当。
Oligonucleotide probe arrays have enabled massively parallel analysis of gene expression levels from a single cDNA sample. Application of microarray technology to analyzing genomic DNA has been stymied by the sequence complexity of the entire human genome. A robust, single base - resolution direct genomic assay would extend the reach of microarray technology. We developed an array-based whole-genome genotyping assay that does not require PCR and enables effectively unlimited multiplexing. The assay achieves a high signal-to-noise ratio by combining specific hybridization of picomolar concentrations of whole genome - amplified DNA to arrayed probes with allele-specific primer extension and signal amplification. As proof of principle, we genotyped several hundred previously characterized SNPs. The conversion rate, call rate and accuracy were comparable to those of high-performance PCR-based genotyping assays.