Programme of Genetics, Scottish Crop Research Institute, Dundee, UK.
Biotechniques. 2011 Mar;50(3):165-74. doi: 10.2144/000113627.
Second-generation sequencing now provides the potential for low-cost generation of whole-genome sequences. However, for large-genome organisms with high repetitive DNA content, genome-wide short read sequence assembly is currently impossible, with accurate ordering and localization of genes still relying heavily on integration with physical and genetic maps. To facilitate this process, we have used Agilent microarrays to simultaneously address thousands of gene sequences to individual BAC clones and contiguous sequences that form part of an emerging physical map of the large and currently unsequenced 5.3-Gb barley genome. The approach represents a cost-effective, highly parallel alternative to traditional addressing methods. By coupling the gene-to-BAC address data with gene-based molecular markers, thousands of BACs can be anchored directly to the genetic map, thereby generating a framework for orientating and ordering genes, and providing direct links to phenotypic traits.
第二代测序现在为低成本生成全基因组序列提供了可能性。然而，对于具有高重复 DNA 含量的大基因组生物来说，目前还不可能对全基因组的短读序列进行组装，而基因的准确排序和定位仍然严重依赖于与物理图谱和遗传图谱的整合。为了促进这一过程，我们使用安捷伦微阵列同时针对数千个基因序列进行了定位，这些基因序列分别位于 BAC 克隆和连续序列上，这些连续序列构成了大型且尚未测序的 5.3Gb 大麦基因组新兴物理图谱的一部分。这种方法代表了一种具有成本效益的、高度并行的替代传统寻址方法。通过将基因到 BAC 的地址数据与基于基因的分子标记相结合，成千上万的 BAC 可以直接锚定到遗传图谱上，从而为基因的定向和排序生成一个框架，并提供与表型特征的直接联系。
