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SSPACE-LongRead:利用长读段序列信息搭建细菌草图基因组支架

SSPACE-LongRead: scaffolding bacterial draft genomes using long read sequence information.

作者信息

Boetzer Marten, Pirovano Walter

机构信息

BaseClear B,V,, Genome analysis and technology department, Einsteinweg 5, Leiden 2333 CC, The Netherlands.

出版信息

BMC Bioinformatics. 2014 Jun 20;15:211. doi: 10.1186/1471-2105-15-211.

Abstract

BACKGROUND

The recent introduction of the Pacific Biosciences RS single molecule sequencing technology has opened new doors to scaffolding genome assemblies in a cost-effective manner. The long read sequence information is promised to enhance the quality of incomplete and inaccurate draft assemblies constructed from Next Generation Sequencing (NGS) data.

RESULTS

Here we propose a novel hybrid assembly methodology that aims to scaffold pre-assembled contigs in an iterative manner using PacBio RS long read information as a backbone. On a test set comprising six bacterial draft genomes, assembled using either a single Illumina MiSeq or Roche 454 library, we show that even a 50× coverage of uncorrected PacBio RS long reads is sufficient to drastically reduce the number of contigs. Comparisons to the AHA scaffolder indicate our strategy is better capable of producing (nearly) complete bacterial genomes.

CONCLUSIONS

The current work describes our SSPACE-LongRead software which is designed to upgrade incomplete draft genomes using single molecule sequences. We conclude that the recent advances of the PacBio sequencing technology and chemistry, in combination with the limited computational resources required to run our program, allow to scaffold genomes in a fast and reliable manner.

摘要

背景

近期太平洋生物科学公司的RS单分子测序技术的引入,为以经济高效的方式构建基因组支架开辟了新途径。长读长序列信息有望提高由下一代测序(NGS)数据构建的不完整和不准确的草图组装的质量。

结果

在此,我们提出了一种新颖的混合组装方法,旨在以迭代方式使用PacBio RS长读长信息作为主干来构建预组装的重叠群。在一个由六个细菌草图基因组组成的测试集上,这些基因组使用单个Illumina MiSeq或罗氏454文库进行组装,我们表明,即使50倍覆盖未校正的PacBio RS长读长也足以大幅减少重叠群的数量。与AHA支架构建器的比较表明,我们的策略更有能力产生(近乎)完整的细菌基因组。

结论

当前工作描述了我们的SSPACE-LongRead软件,其设计目的是使用单分子序列升级不完整的草图基因组。我们得出结论,PacBio测序技术和化学的最新进展,结合运行我们程序所需的有限计算资源,能够以快速可靠的方式构建基因组支架。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d224/4076250/dd6af88d13e1/1471-2105-15-211-1.jpg

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