UBC Faculty Research and Publications

Sequencing platform and library preparation choices impact viral metagenomes Solonenko, Sergei A; Ignacio-Espinoza, J C; Alberti, Adriana; Cruaud, Corinne; Hallam, Steven; Konstantinidis, Kostas; Tyson, Gene; Wincker, Patrick; Sullivan, Matthew B

Abstract

Background: Microbes drive the biogeochemistry that fuels the planet. Microbial viruses modulate their hosts directly through mortality and horizontal gene transfer, and indirectly by re-programming host metabolisms during infection. However, our ability to study these virus-host interactions is limited by methods that are low-throughput and heavily reliant upon the subset of organisms that are in culture. One way forward are culture-independent metagenomic approaches, but these novel methods are rarely rigorously tested, especially for studies of environmental viruses, air microbiomes, extreme environment microbiology and other areas with constrained sample amounts. Here we perform replicated experiments to evaluate Roche 454, Illumina HiSeq, and Ion Torrent PGM sequencing and library preparation protocols on virus metagenomes generated from as little as 10pg of DNA. Results Using %G + C content to compare metagenomes, we find that (i) metagenomes are highly replicable, (ii) some treatment effects are minimal, e.g., sequencing technology choice has 6-fold less impact than varying input DNA amount, and (iii) when restricted to a limited DNA concentration (

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Attribution 4.0 International (CC BY 4.0)