Department of Molecular, Cellular, and Developmental Biology, University of California Santa Cruz, Santa Cruz, CA 95064, USA.
Department of Biomolecular Engineering, University of California Santa Cruz, Santa Cruz, CA 95064, USA.
Nucleic Acids Res. 2020 Jul 27;48(13):e75. doi: 10.1093/nar/gkaa450.
A high quality genome assembly is a vital first step for the study of an organism. Recent advances in technology have made the creation of high quality chromosome scale assemblies feasible and low cost. However, the amount of input DNA needed for an assembly project can be a limiting factor for small organisms or precious samples. Here we demonstrate the feasibility of creating a chromosome scale assembly using a hybrid method for a low input sample, a single outbred Drosophila melanogaster. Our approach combines an Illumina shotgun library, Oxford nanopore long reads, and chromosome conformation capture for long range scaffolding. This single fly genome assembly has a N50 of 26 Mb, a length that encompasses entire chromosome arms, contains 95% of expected single copy orthologs, and a nearly complete assembly of this individual's Wolbachia endosymbiont. The methods described here enable the accurate and complete assembly of genomes from small, field collected organisms as well as precious clinical samples.
高质量的基因组组装是研究生物的重要第一步。最近技术的进步使得高质量的染色体水平组装变得可行且成本低廉。然而,对于小型生物或珍贵样本来说,组装项目所需的输入 DNA 量可能是一个限制因素。在这里,我们展示了使用混合方法从小量样本(一只单交果蝇)中创建染色体水平组装的可行性。我们的方法结合了 Illumina shotgun 文库、Oxford nanopore 长读长和染色体构象捕获技术进行长距离支架构建。这个单只果蝇基因组组装的 N50 为 26Mb,长度涵盖了整个染色体臂,包含了 95%预期的单拷贝直系同源物,并且几乎完整地组装了该个体的沃尔巴克氏体共生菌。这里描述的方法可用于从小型、野外采集的生物以及珍贵的临床样本中准确、完整地组装基因组。