Jupe Florian, Chen Xinwei, Verweij Walter, Witek Kamel, Jones Jonathan D G, Hein Ingo
The Sainsbury Laboratory, Norwich Research Park, Norwich, NR4 7UH, UK.
Methods Mol Biol. 2014;1127:291-303. doi: 10.1007/978-1-62703-986-4_22.
Enrichment of genomic DNA for genes of interest prior to high-throughput sequencing offers an efficient and cost-effective approach to reduce genome complexity. Target enrichment typically yields higher read-depth for selected genes and is therefore suitable for determination of sequence polymorphisms and enables multiplexing of samples. Target enrichment also provides a means to annotate specific gene families within the sequenced organisms without the requirements for gene models. Here we describe enrichment procedures for NB-LRR-type plant resistance genes that can, for example, be used to establish the NB-LRR gene complements of individual plants and to map resistances more rapidly using a bulked segregant analysis.
在高通量测序之前对感兴趣的基因进行基因组DNA富集,为降低基因组复杂性提供了一种高效且经济有效的方法。靶向富集通常会为选定的基因产生更高的读取深度,因此适用于确定序列多态性,并能实现样本的多重分析。靶向富集还提供了一种方法,可在无需基因模型的情况下注释测序生物体内的特定基因家族。在此,我们描述了NB-LRR型植物抗性基因的富集程序,例如,该程序可用于确定单个植物的NB-LRR基因互补情况,并通过混合分离群体分析更快地定位抗性。