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Identifying contamination with advanced visualization and analysis practices: metagenomic approaches for eukaryotic genome assemblies

机译:通过先进的可视化和分析方法识别污染:真核生物基因组组装的宏基因组学方法

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摘要

High-throughput sequencing provides a fast and cost-effective mean to recover genomes of organisms from all domains of life. However, adequate curation of the assembly results against potential contamination of non-target organisms requires advanced bioinformatics approaches and practices. Here, we re-analyzed the sequencing data generated for the tardigrade Hypsibius dujardini, and created a holistic display of the eukaryotic genome assembly using DNA data originating from two groups and eleven sequencing libraries. By using bacterial single-copy genes, k-mer frequencies, and coverage values of scaffolds we could identify and characterize multiple near-complete bacterial genomes from the raw assembly, and curate a 182 Mbp draft genome for H. dujardini supported by RNA-Seq data. Our results indicate that most contaminant scaffolds were assembled from Moleculo long-read libraries, and most of these contaminants have differed between library preparations. Our re-analysis shows that visualization and curation of eukaryotic genome assemblies can benefit from tools designed to address the needs of today’s microbiologists, who are constantly challenged by the difficulties associated with the identification of distinct microbial genomes in complex environmental metagenomes.
机译:高通量测序提供了一种快速且经济高效的手段,可从生命的所有领域中恢复生物的基因组。但是,对组装结果进行适当的处​​理以防止对非目标生物的潜在污染,需要先进的生物信息学方法和实践。在这里,我们重新分析了为缓节菌Hypsibius dujardini生成的测序数据,并使用源自两组和11个测序文库的DNA数据创建了真核生物基因组装配的整体展示。通过使用细菌单拷贝基因,k-mer频率和支架的覆盖值,我们可以从原始装配中鉴定和表征多个接近完整的细菌基因组,并为RNA-Seq支持的杜氏jar草拟定182 Mbp的基因组草案数据。我们的结果表明,大多数污染物支架是从Moleculo长读文库中组装的,并且这些污染物中的大多数在文库制备之间存在差异。我们的重新分析表明,真核生物基因组组装的可视化和管理可从旨在满足当今微生物学家需求的工具中受益,这些微生物学家经常面临与复杂环境元基因组中不同微生物基因组鉴定相关的困难。

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