The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.
Cancer Research UK Cambridge Institute, Li Ka Shing Centre, University of Cambridge, Cambridge, CB2 0RE, UK.
Sci Rep. 2017 Jun 21;7(1):3935. doi: 10.1038/s41598-017-03996-z.
Long-read sequencing technologies such as Pacific Biosciences and Oxford Nanopore MinION are capable of producing long sequencing reads with average fragment lengths of over 10,000 base-pairs and maximum lengths reaching 100,000 base- pairs. Compared with short reads, the assemblies obtained from long-read sequencing platforms have much higher contig continuity and genome completeness as long fragments are able to extend paths into problematic or repetitive regions. Many successful assembly applications of the Pacific Biosciences technology have been reported ranging from small bacterial genomes to large plant and animal genomes. Recently, genome assemblies using Oxford Nanopore MinION data have attracted much attention due to the portability and low cost of this novel sequencing instrument. In this paper, we re-sequenced a well characterized genome, the Saccharomyces cerevisiae S288C strain using three different platforms: MinION, PacBio and MiSeq. We present a comprehensive metric comparison of assemblies generated by various pipelines and discuss how the platform associated data characteristics affect the assembly quality. With a given read depth of 31X, the assemblies from both Pacific Biosciences and Oxford Nanopore MinION show excellent continuity and completeness for the 16 nuclear chromosomes, but not for the mitochondrial genome, whose reconstruction still represents a significant challenge.
长读测序技术,如 Pacific Biosciences 和 Oxford Nanopore MinION,能够产生平均片段长度超过 10000 个碱基对、最长可达 100000 个碱基对的长测序读段。与短读段相比,长读段测序平台获得的组装具有更高的连续度和基因组完整性,因为长片段能够延伸到有问题或重复的区域。Pacific Biosciences 技术的许多成功组装应用已经被报道,从小细菌基因组到大型植物和动物基因组。最近,由于这种新型测序仪器的便携性和低成本,使用 Oxford Nanopore MinION 数据的基因组组装引起了广泛关注。在本文中,我们使用三种不同的平台:MinION、PacBio 和 MiSeq,重新测序了一个特征良好的基因组,酿酒酵母 S288C 株。我们对来自不同管道的组装结果进行了全面的指标比较,并讨论了平台相关数据特征如何影响组装质量。在给定的 31X 读深度下,来自 Pacific Biosciences 和 Oxford Nanopore MinION 的组装对 16 条核染色体表现出极好的连续性和完整性,但对线粒体基因组却不行,其重建仍然是一个重大挑战。