Data from: Quantitative DNA metabarcoding: improved estimates of species proportional biomass using correction factors derived from control material

Dryad version number: 1 Version status: submitted Dryad curation status: Published Sharing link: https://datadryad.org/stash/share/pO28HGR3P868SmT7zbgGs663nfkkogrQfWNeZGF01aw Storage size: 745061847 Visibility: public Usage notes Model System Data Please see ReadMe file Prey_Library_Data Please see...

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Bibliographic Details
Main Authors: Thomas, Austen C., Deagle, Bruce E., Eveson, J. Paige, Harsch, Corie H., Trites, Andrew W.
Other Authors: Federated Research Data Repository, Dépôt fédéré de données de recherche
Format: Dataset
Language:unknown
Published: Dryad Digital Repository 2015
Subjects:
Online Access:https://doi.org/10.5061/dryad.7dv96
https://doi.org/10.5683/sp2/l3d0ru
https://doi.org/10.14288/1.0398084
Description
Summary:Dryad version number: 1 Version status: submitted Dryad curation status: Published Sharing link: https://datadryad.org/stash/share/pO28HGR3P868SmT7zbgGs663nfkkogrQfWNeZGF01aw Storage size: 745061847 Visibility: public Usage notes Model System Data Please see ReadMe file Prey_Library_Data Please see ReadMe file Supplemental_Files Tissue mix sequence counts Custom BLAST database Model system species sequences Abstract DNA metabarcoding is a powerful new tool allowing characterization of species assemblages using high-throughput amplicon sequencing. The utility of DNA metabarcoding for quantifying relative species abundances is currently limited by both biological and technical biases which influence sequence read counts. We tested the idea of sequencing 50/50 mixtures of target species and a control species in order to generate relative correction factors (RCFs) that account for multiple sources of bias and are applicable to field studies. RCFs will be most effective if they are not affected by input mass ratio or co-occurring species. In a model experiment involving three target fish species and a fixed control, we found RCFs did vary with input ratio but in a consistent fashion, and that 50/50 RCFs applied to DNA sequence counts from various mixtures of the target species still greatly improved relative abundance estimates (e.g. average per species error of 19 ± 8% for uncorrected vs. 3 ± 1% for corrected estimates). To demonstrate the use of correction factors in a field setting, we calculated 50/50 RCFs for 18 harbour seal (Phoca vitulina) prey species (RCFs ranging from 0.68 to 3.68). Applying these corrections to field-collected seal scats affected species percentages from individual samples (Δ 6.7 ± 6.6%) more than population-level species estimates (Δ 1.7 ± 1.2%). Our results indicate that the 50/50 RCF approach is an effective tool for evaluating and correcting biases in DNA metabarcoding studies. The decision to apply correction factors will be influenced by the feasibility of creating tissue mixtures ...