Robust high-throughput prokaryote de novo assembly and improvement pipeline for Illumina data

• Published in: Microbial genomics Vol. 2; no. 8; p. e000083
• Main Authors: Page, Andrew J., De Silva, Nishadi, Hunt, Martin, Quail, Michael A., Parkhill, Julian, Harris, Simon R., Otto, Thomas D., Keane, Jacqueline A.
• Format: Journal Article
• Language: English
• Published: England Microbiology Society 01.08.2016
• Subjects: Genome, Bacterial - genetics; Genomics - economics; Genomics - methods; High-Throughput Nucleotide Sequencing; Methods Paper; Multilocus Sequence Typing; Prokaryotic Cells; Sequence Analysis, DNA - methods; Software; illumina; assembly; high-throughput; prokaryotic
• ISSN: 2057-5858; 2057-5858
• DOI: 10.1099/mgen.0.000083

The rapidly reducing cost of bacterial genome sequencing has lead to its routine use in large-scale microbial analysis. Though mapping approaches can be used to find differences relative to the reference, many bacteria are subject to constant evolutionary pressures resulting in events such as the loss and gain of mobile genetic elements, horizontal gene transfer through recombination and genomic rearrangements. De novo assembly is the reconstruction of the underlying genome sequence, an essential step to understanding bacterial genome diversity. Here we present a high-throughput bacterial assembly and improvement pipeline that has been used to generate nearly 20 000 annotated draft genome assemblies in public databases. We demonstrate its performance on a public data set of 9404 genomes. We find all the genes used in multi-locus sequence typing schema present in 99.6 % of assembled genomes. When tested on low-, neutral- and high-GC organisms, more than 94 % of genes were present and completely intact. The pipeline has been proven to be scalable and robust with a wide variety of datasets without requiring human intervention. All of the software is available on GitHub under the GNU GPL open source license.