Quantitative DNA metabarcoding: improved estimates of species proportional biomass using correction factors derived from control material
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...
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ftunivtasecite:oai:ecite.utas.edu.au:118434 2023-05-15T16:33:39+02:00 Quantitative DNA metabarcoding: improved estimates of species proportional biomass using correction factors derived from control material Thomas, AC Deagle, BE Eveson, JP Harsch, CH Trites, AW 2016 https://doi.org/10.1111/1755-0998.12490 http://www.ncbi.nlm.nih.gov/pubmed/26602877 http://ecite.utas.edu.au/118434 en eng Wiley-Blackwell Publishing Ltd. http://dx.doi.org/10.1111/1755-0998.12490 Thomas, AC and Deagle, BE and Eveson, JP and Harsch, CH and Trites, AW, Quantitative DNA metabarcoding: improved estimates of species proportional biomass using correction factors derived from control material, Molecular Ecology Resources, 16, (3) pp. 714-726. ISSN 1755-098X (2016) [Refereed Article] http://www.ncbi.nlm.nih.gov/pubmed/26602877 http://ecite.utas.edu.au/118434 Biological Sciences Genetics Genetics not elsewhere classified Refereed Article PeerReviewed 2016 ftunivtasecite https://doi.org/10.1111/1755-0998.12490 2022-08-22T22:16:50Z 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 198% for uncorrected vs. 31% 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 6.76.6%) more than population-level species estimates (Δ 1.71.2%). 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 for the target species, and the level of accuracy needed to meet research objectives. Article in Journal/Newspaper harbour seal Phoca vitulina eCite UTAS (University of Tasmania) Molecular Ecology Resources 16 3 714 726 |
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English |
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Biological Sciences Genetics Genetics not elsewhere classified |
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Biological Sciences Genetics Genetics not elsewhere classified Thomas, AC Deagle, BE Eveson, JP Harsch, CH Trites, AW Quantitative DNA metabarcoding: improved estimates of species proportional biomass using correction factors derived from control material |
topic_facet |
Biological Sciences Genetics Genetics not elsewhere classified |
description |
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 198% for uncorrected vs. 31% 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 6.76.6%) more than population-level species estimates (Δ 1.71.2%). 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 for the target species, and the level of accuracy needed to meet research objectives. |
format |
Article in Journal/Newspaper |
author |
Thomas, AC Deagle, BE Eveson, JP Harsch, CH Trites, AW |
author_facet |
Thomas, AC Deagle, BE Eveson, JP Harsch, CH Trites, AW |
author_sort |
Thomas, AC |
title |
Quantitative DNA metabarcoding: improved estimates of species proportional biomass using correction factors derived from control material |
title_short |
Quantitative DNA metabarcoding: improved estimates of species proportional biomass using correction factors derived from control material |
title_full |
Quantitative DNA metabarcoding: improved estimates of species proportional biomass using correction factors derived from control material |
title_fullStr |
Quantitative DNA metabarcoding: improved estimates of species proportional biomass using correction factors derived from control material |
title_full_unstemmed |
Quantitative DNA metabarcoding: improved estimates of species proportional biomass using correction factors derived from control material |
title_sort |
quantitative dna metabarcoding: improved estimates of species proportional biomass using correction factors derived from control material |
publisher |
Wiley-Blackwell Publishing Ltd. |
publishDate |
2016 |
url |
https://doi.org/10.1111/1755-0998.12490 http://www.ncbi.nlm.nih.gov/pubmed/26602877 http://ecite.utas.edu.au/118434 |
genre |
harbour seal Phoca vitulina |
genre_facet |
harbour seal Phoca vitulina |
op_relation |
http://dx.doi.org/10.1111/1755-0998.12490 Thomas, AC and Deagle, BE and Eveson, JP and Harsch, CH and Trites, AW, Quantitative DNA metabarcoding: improved estimates of species proportional biomass using correction factors derived from control material, Molecular Ecology Resources, 16, (3) pp. 714-726. ISSN 1755-098X (2016) [Refereed Article] http://www.ncbi.nlm.nih.gov/pubmed/26602877 http://ecite.utas.edu.au/118434 |
op_doi |
https://doi.org/10.1111/1755-0998.12490 |
container_title |
Molecular Ecology Resources |
container_volume |
16 |
container_issue |
3 |
container_start_page |
714 |
op_container_end_page |
726 |
_version_ |
1766023335599144960 |