Abstract

BackgroundAnalysis of mixed microbial communities using metagenomic sequencing experiments requires multiple preprocessing and analytical steps to interpret the microbial and genetic composition of samples. Analytical steps include quality control, adapter trimming, host decontamination, metagenomic classification, read assembly, and alignment to reference genomes.ResultsWe present a modular and user-extensible pipeline called Sunbeam that performs these steps in a consistent and reproducible fashion. It can be installed in a single step, does not require administrative access to the host computer system, and can work with most cluster computing frameworks. We also introduce Komplexity, a software tool to eliminate potentially problematic, low-complexity nucleotide sequences from metagenomic data. A unique component of the Sunbeam pipeline is an easy-to-use extension framework that enables users to add custom processing or analysis steps directly to the workflow. The pipeline and its extension framework are well documented, in routine use, and regularly updated.ConclusionsSunbeam provides a foundation to build more in-depth analyses and to enable comparisons in metagenomic sequencing experiments by removing problematic, low-complexity reads and standardizing post-processing and analytical steps. Sunbeam is written in Python using the Snakemake workflow management software and is freely available at github.com/sunbeam-labs/sunbeam under the GPLv3.

Highlights

  • Analysis of mixed microbial communities using metagenomic sequencing experiments requires multiple preprocessing and analytical steps to interpret the microbial and genetic composition of samples

  • The capabilities of Sunbeam compare favorably with features offered in existing pipelines such as SURPI (Sequence-based Ultra-Rapid Pathogen Identification) [26], EDGE (Empowering the Development of Genomics Expertise) [27], ATLAS (Automatic Tool for Local Assembly Structures) [28], and KneadData [29]

  • The Sunbeam extensions framework facilitates the addition of new features to Sunbeam without adding overhead to the core pipeline—once developed, a Sunbeam extension can be discovered and used by anyone through our extensions website

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Summary

Introduction

Analysis of mixed microbial communities using metagenomic sequencing experiments requires multiple preprocessing and analytical steps to interpret the microbial and genetic composition of samples. Metagenomic shotgun sequencing involves isolating DNA from a mixed microbial community of interest, sequencing deeply into DNAs drawn randomly from the mixture. This is in contrast to marker gene sequencing (e.g., the 16S rRNA gene of bacteria), where a specific target gene region is amplified and sequenced. Tools, or reference database versions between analyses makes it challenging to compare the results of different metagenomic sequencing experiments (2019) 7:46

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