Kilian Andrzej, Wenzl Peter, Huttner Eric, Carling Jason, Xia Ling, Blois Hélène, Caig Vanessa, Heller-Uszynska Katarzyna, Jaccoud Damian, Hopper Colleen, Aschenbrenner-Kilian Malgorzata, Evers Margaret, Peng Kaiman, Cayla Cyril, Hok Puthick, Uszynski Grzegorz
Diversity Arrays Technology Pty Ltd, Yarralumla, Canberra, ACT, Australia.
Methods Mol Biol. 2012;888:67-89. doi: 10.1007/978-1-61779-870-2_5.
In the last 20 years, we have observed an exponential growth of the DNA sequence data and simular increase in the volume of DNA polymorphism data generated by numerous molecular marker technologies. Most of the investment, and therefore progress, concentrated on human genome and genomes of selected model species. Diversity Arrays Technology (DArT), developed over a decade ago, was among the first "democratizing" genotyping technologies, as its performance was primarily driven by the level of DNA sequence variation in the species rather than by the level of financial investment. DArT also proved more robust to genome size and ploidy-level differences among approximately 60 organisms for which DArT was developed to date compared to other high-throughput genotyping technologies. The success of DArT in a number of organisms, including a wide range of "orphan crops," can be attributed to the simplicity of underlying concepts: DArT combines genome complexity reduction methods enriching for genic regions with a highly parallel assay readout on a number of "open-access" microarray platforms. The quantitative nature of the assay enabled a number of applications in which allelic frequencies can be estimated from DArT arrays. A typical DArT assay tests for polymorphism tens of thousands of genomic loci with the final number of markers reported (hundreds to thousands) reflecting the level of DNA sequence variation in the tested loci. Detailed DArT methods, protocols, and a range of their application examples as well as DArT's evolution path are presented.
在过去20年里，我们观察到DNA序列数据呈指数增长，并且通过众多分子标记技术生成的DNA多态性数据量也有类似增长。大部分投资以及由此取得的进展都集中在人类基因组和选定模式物种的基因组上。十多年前开发的多样性阵列技术（DArT）是首批“平民化”基因分型技术之一，因为其性能主要由物种中的DNA序列变异水平驱动，而非由资金投入水平驱动。与其他高通量基因分型技术相比，对于迄今已开发出DArT技术的约60种生物，DArT在基因组大小和倍性水平差异方面也表现得更为稳健。DArT在包括多种“小众作物”在内的许多生物中取得成功，可归因于其基本概念的简单性：DArT将富集基因区域的基因组复杂性降低方法与在多个“开放获取”微阵列平台上的高度平行检测读数相结合。该检测的定量性质使得能够进行许多应用，其中等位基因频率可从DArT阵列中估计出来。典型的DArT检测会测试数万个基因组位点的多态性，最终报告的标记数量（数百到数千个）反映了测试位点的DNA序列变异水平。本文介绍了详细的DArT方法、方案及其一系列应用示例以及DArT的发展历程。
