NAMERS: a purpose-built reference DNA sequence database to support applied eDNA metabarcoding
Authors/Creators
- 1. Fisheries and Oceans Canada, Nanaimo, Canada
- 2. Centre for Environmental Genomics Applications/eDNA tech, St. John's, Canada
- 3. University of Guelph, Guelph, Canada|Centre for Environmental Genomics Applications/eDNA tech, St. John's, Canada
Description
Applied eDNA metabarcoding is increasingly being considered as a tool to inform management decisions, regulations, or policy development. Because these downstream considerations are coming to the forefront of eDNA applications, optimizing workflow elements is essential to increasing standardization, efficiency, and competency of metabarcoding results. Reference DNA sequences are critical workflow elements that currently lack consistent approaches to generating, curating, or publishing. We present a complete mitochondrial genome and nuclear ribosomal DNA cistron reference DNA sequence library for 92% of the freshwater fish species of British Columbia, Canada. This resource is published as the Novel Applied eDNA Metabarcoding Reference Sequences (NAMERS) repository (https://namers.ca), a user-friendly and interactive website for specialists and non-specialists alike to explore and generate custom reference libraries for taxa and genes of interest. We demonstrate the power of NAMERS for optimization of applied eDNA metabarcoding study design by analyzing the number of primer mismatches and species resolution power of existing metabarcoding markers. NAMERS demonstrates that high quality curated genomic information is within a reasonable reach to meet the increasing demand for actionable eDNA metabarcoding applications. The framework used here incorporating the pillars of accuracy, completeness and accessibility can be applied for new iterations of other reference sequence databases to bring DNA-based monitoring into a new era.
Files
MBMG_article_125095.pdf
Files
(2.1 MB)
| Name | Size | Download all |
|---|---|---|
|
md5:8d322c89eb747e075a148c65010e8f76
|
2.1 MB | Preview Download |
System files
(171.5 kB)
| Name | Size | Download all |
|---|---|---|
|
md5:cdd53c654e5fb8c4e22137ca76964a97
|
171.5 kB | Download |
Linked records
Additional details
References
- Allison MJ, Warren RL, Lopez ML, Acharya-Patel N, Imbery JJ, Coombe L, Yang CL, Birol I, Helbing CC (2023) Enabling robust environmental DNA assay design with "unikseq" for the identification of taxon-specific regions within whole mitochondrial genomes. Environmental DNA 5(5): 1032–1047. https://doi.org/10.1002/edn3.438
- Atkas C (2020) haplotypes: Manipulating DNA Sequences and Estimating Unambiguous Haplotype Network with Statistical Parsimony. https://cran.r-project.org/web/packages/haplotypes/index.html
- Baird DJ, Hajibabaei M (2012) Biomonitoring 2.0: A new paradigm in ecosystem assessment made possible by next-generation DNA sequencing. Molecular Ecology 21(8): 2039–2044. https://doi.org/10.1111/j.1365-294X.2012.05519.x
- Beamish RJ (1987) Evidence that parasitic and non-parasitic life history types are produced by one population of lamprey. Canadian Journal of Fisheries and Aquatic Sciences 44(10): 1779–1782. https://doi.org/10.1139/f87-219
- Benson DA, Karsch-Mizrachi I, Lipman DJ, Ostell J, Rapp BA, Wheeler DL (2000) GenBank. Nucleic Acids Research 28(1): 15–18. https://doi.org/10.1093/nar/28.1.15
- Berry TE, Osterrieder SK, Murray DC, Coghlan ML, Richardson AJ, Grealy AK, Stat M, Bejder L, Bunce M (2017) DNA metabarcoding for diet analysis and biodiversity: A case study using the endangered Australian sea lion (Neophoca cinerea). Ecology and Evolution 7(14): 5435–5453. https://doi.org/10.1002/ece3.3123
- Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: A flexible trimmer for Illumina sequence data. Bioinformatics (Oxford, England) 30(15): 2114–2120. https://doi.org/10.1093/bioinformatics/btu170
- Bylemans J, Gleeson DM, Hardy CM, Furlan E (2018) Toward an ecoregion scale evaluation of eDNA metabarcoding primers: A case study for the freshwater fish biodiversity of the Murray–Darling Basin (Australia). Ecology and Evolution 8(17): 8697–8712. https://doi.org/10.1002/ece3.4387
- Claver C, Canals O, de Amézaga LG, Mendibil I, Rodriguez-Ezpeleta N (2023) An automated workflow to assess completeness and curate GenBank for environmental DNA metabarcoding: The marine fish assemblage as case study. Environmental DNA 5(4): 634–647. https://doi.org/10.1002/edn3.433
- Collins RA, Bakker J, Wangensteen OS, Soto AZ, Corrigan L, Sims DW, Genner MJ, Mariani S (2019) Non-specific amplification compromises environmental DNA metabarcoding with COI. Methods in Ecology and Evolution 10(11): 1985–2001. https://doi.org/10.1111/2041-210X.13276
- Collins RA, Trauzzi G, Maltby KM, Gibson TI, Ratcliffe FC, Hallam J, Rainbird S, Maclaine J, Henderson PA, Sims DW, Mariani S, Genner MJ (2021) Meta-Fish-Lib: A generalised, dynamic DNA reference library pipeline for metabarcoding of fishes. Journal of Fish Biology 99(4): 1446–1454. https://doi.org/10.1111/jfb.14852
- Cordier T, Alonso-Sáez L, Apothéloz-Perret-Gentil L, Aylagas E, Bohan DA, Bouchez A, Chariton A, Creer S, Frühe L, Keck F, Keeley N, Laroche O, Leese F, Pochon X, Stoeck T, Pawlowski J, Lanzén A (2021) Ecosystems monitoring powered by environmental genomics: A review of current strategies with an implementation roadmap. Molecular Ecology 30(13): 2937–2958. https://doi.org/10.1111/mec.15472
- Creer S, Deiner K, Frey S, Porazinska D, Taberlet P, Thomas WK, Potter C, Bik HM (2016) The ecologist's field guide to sequence-based identification of biodiversity. Methods in Ecology and Evolution 7(9): 1008–1018. https://doi.org/10.1111/2041-210X.12574
- Cristescu ME, Hebert PDN (2018) Uses and misuses of environmental DNA in biodiversity science and conservation. Annual Review of Ecology, Evolution, and Systematics 49(1): 209–230. https://doi.org/10.1146/annurev-ecolsys-110617-062306
- Darling JA, Mahon AR (2011) From molecules to management: Adopting DNA-based methods for monitoring biological invasions in aquatic environments. Environmental Research 111(7): 978–988. https://doi.org/10.1016/j.envres.2011.02.001
- Darling JA, Pochon X, Abbott CL, Inglis GJ, Zaiko A (2020) The risks of using molecular biodiversity data for incidental detection of species of concern. Diversity & Distributions 26(9): 1116–1121. https://doi.org/10.1111/ddi.13108
- Darling JA, Jerde CL, Sepulveda AJ (2021) What do you mean by false positive? Environmental DNA 3(5): 879–883. https://doi.org/10.1002/edn3.194
- Deagle BE, Jarman SN, Coissac E, Pompanon F, Taberlet P (2014) DNA metabarcoding and the cytochrome c oxidase subunit I marker: Not a perfect match. Biology Letters 10(9): 20140562. https://doi.org/10.1098/rsbl.2014.0562
- Dziedzic E, Sidlauskas B, Cronn R, Anthony J, Cornwell T, Friesen TA, Konstantinidis P, Penaluna BE, Stein S, Levi T (2023) Creating, curating and evaluating a mitogenomic reference database to improve regional species identification using environmental DNA. Molecular Ecology Resources 23(8): 1880–1904. https://doi.org/10.1111/1755-0998.13855
- Edgar RC (2004) MUSCLE: Multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research 32(5): 1792–1797. https://doi.org/10.1093/nar/gkh340
- European Parliament & Council of the European Union (2008) Directive establishing a framework for community action in the field of marine environmental policy (Marine Strategy Framework Directive). Directive 2008/56/EC. European Commission. Joint Research Centre. Institute for Environment and Sustainability. Marine Strategy Framework Directive - Competence Centre. https://mcc.jrc.ec.europa.eu/main/index.py
- Evans NT, Olds BP, Renshaw MA, Turner CR, Li Y, Jerde CL, Mahon AR, Pfrender ME, Lamberti GA, Lodge DM (2016) Quantification of mesocosm fish and amphibian species diversity via environmental DNA metabarcoding. Molecular Ecology Resources 16(1): 29–41. https://doi.org/10.1111/1755-0998.12433
- Feist SM, Lance RF (2021) Advanced molecular-based surveillance of quagga and zebra mussels: A review of environmental DNA/RNA (eDNA/eRNA) studies and considerations for future directions. NeoBiota 66: 117–159. https://doi.org/10.3897/neobiota.66.60751
- Ficetola GF, Boyer F, Valentini A, Bonin A, Meyer A, Dejean T, Gaboriaud C, Usseglio-Polatera P, Taberlet P (2021) Comparison of markers for the monitoring of freshwater benthic biodiversity through DNA metabarcoding. Molecular Ecology 30(13): 3189–3202. https://doi.org/10.1111/mec.15632
- Gold Z, Curd EE, Goodwin KD, Choi ES, Frable BW, Thompson AR, Walker Jr HJ, Burton RS, Kacev D, Martz LD, Barber PH (2021) Improving metabarcoding taxonomic assignment: A case study of fishes in a large marine ecosystem. Molecular Ecology Resources 21(7): 2546–2564. https://doi.org/10.1111/1755-0998.13450
- Goodwin KD, Aiello CM, Weise M, Edmondson M, Fillingham K, Allen D, Amerson A, Barton ML, Benson A, Canonico G, Gold Z, Gumm J, Hunter M, Joffe N, Lance R, Larkin A, Letelier R, Lipsky C, McCoskey D, Morrison C, Clark K, Darling JA, Demery A-J, Everett M, Fletcher-Hoppe C, Nichols KM, Parsons KM, Price J, Puglise K, Scholl K, Schwartz MK, Sepulveda A, Shannon J, Turner W, White T (2024) National Aquatic Environmental DNA Strategy. US Geological Survey Report, Wetland and Aquatic Research Centre, 70255545. https://www.usgs.gov/publications/national-aquatic-environmental-dna-strategy
- Hajibabaei M (2022) Demystifying eDNA validation. Trends in Ecology & Evolution 37(10): 826–828. https://doi.org/10.1016/j.tree.2022.06.015
- Hajibabaei M, Shokralla S, Zhou X, Singer GAC, Baird DJ (2011) Environmental barcoding: A next-generation sequencing approach for biomonitoring applications using river benthos. PLOS ONE 6(4): e17497. https://doi.org/10.1371/journal.pone.0017497
- Hering D, Borja A, Jones JI, Pont D, Boets P, Bouchez A, Bruce K, Drakare S, Hanfling B, Kahlert M, Leese F, Meissner K, Mergen P, Reyjol Y, Segurado P, Vogler A, Kelly M (2018) Implementation options for DNA-based identification into ecological status assessment under the European Water Framework Directive. Water Research 138: 192–205. https://doi.org/10.1016/j.watres.2018.03.003
- Hoban ML, Whitney J, Collins AG, Meyer C, Murphy KR, Reft AJ, Bemis KE (2022) Skimming for barcodes: Rapid production of mitochondrial genome and nuclear ribosomal repeat reference markers through shallow shotgun sequencing. PeerJ 10: e13790. https://doi.org/10.7717/peerj.13790
- Hubert N, Hanner R, Holm E, Mandrak NE, Taylor E, Burridge M, Watkinson D, Dumont P, Curry A, Bentzen P, Zhang J, April J, Bernatchez L (2008) Identifying Canadian freshwater fishes through DNA barcodes. PLOS ONE 3(6): e2490. https://doi.org/10.1371/journal.pone.0002490
- IBPES (2019) Summary for Policymakers of the Global Assessment Report on Biodiversity and Ecosystem Services of the Intergovernmental Science-Policy Platform on Biodiversity and Ecosystem Services.
- Jin JJ, Yu WB, Yang JB, Song Y, dePamphilis CW, Yi TS, Li DZ (2020) GetOrganelle: A fast and versatile toolkit for accurate de novo assembly of organelle genomes. Genome Biology 21(1): 241. https://doi.org/10.1186/s13059-020-02154-5
- Kelly RP, Lodge DM, Lee KN, Theroux S, Sepulveda AJ, Scholin CA, Craine JM, Andruszkiewicz Allan E, Nichols KM, Parsons KM, Goodwin KD, Gold Z, Chavez FP, Noble RT, Abbott CL, Baerwald MR, Naaum AM, Thielen PM, Simons AL, Jerde CL, Duda JJ, Hunter ME, Hagan JA, Meyer RS, Steele JA, Stoeckle MY, Bik HM, Meyer CP, Stein E, James KE, Thomas AC, Demir-Hilton E, Timmers MA, Griffith JF, Weise MJ, Weisberg SB (2024) Toward a national eDNA strategy for the United States. Environmental DNA 6: e432. https://doi.org/10.1002/edn3.432
- Kitano T, Umetsu K, Tian W, Osawa M (2007) Two universal primer sets for species identification among vertebrates. International Journal of Legal Medicine 121(5): 423–427. https://doi.org/10.1007/s00414-006-0113-y
- Leese F, Bouchez A, Abarenkov K, Altermatt F, Borja A, Bruce K, Ekrem T, Čiampor F, Čiamporová-Zaťovičová Z, Costa FO, Duarte S, Elbrecht V, Fontaneto D, Franc A, Geiger MF, Hering D, Kahlert M, Stroil BK, Kelly M, Keskin E, Liska I, Mergen P, Meissner K, Pawlowski J, Penev L, Reyjol Y, Rotter A, Steinke D, van der Wal B, Vitecek S, Zimmermann J, Weigand AM (2018) Chapter Two - Why We Need Sustainable Networks Bridging Countries, Disciplines, Cultures and Generations for Aquatic Biomonitoring 2.0: A Perspective Derived From the DNAqua-Net COST Action. In: Bohan DA, Dumbrell AJ, Woodward G, Jackson M (Eds) Advances in Ecological Research 58, 63–99. https://doi.org/10.1016/bs.aecr.2018.01.001
- Leray M, Knowlton N, Machida RJ (2022) MIDORI2: A collection of quality controlled, preformatted, and regularly updated reference databases for taxonomic assignment of eukaryotic mitochondrial sequences. Environmental DNA 4(4): 894–907. https://doi.org/10.1002/edn3.303
- Lewin HA, Robinson GE, Kress WJ, Baker WJ, Coddington J, Crandall KA, Durbin R, Edwards SV, Forest F, Gilbert MTP, Goldstein MM, Grigoriev IJ, Hackett KJ, Haussler D, Jarvis ED, Johnson WE, Patrinos A, Richards S, Castilla-Rubio JC, van Sluys M-A, Soltis PS, Xu X, Yang H, Zhang G (2018) Earth BioGenome Project: Sequencing life for the future of life. Proceedings of the National Academy of Sciences of the United States of America 115(17): 4325–4333. https://doi.org/10.1073/pnas.1720115115
- Li JY, Li WX, Wang AT, Zhang Y (2021) MitoFlex: An efficient, high-performance toolkit for animal mitogenome assembly, annotation and visualization. Bioinformatics (Oxford, England) 37(18): 3001–3003. https://doi.org/10.1093/bioinformatics/btab111
- Mathieu C, Hermans SM, Lear G, Buckley TR, Lee KC, Buckley HL (2020) A Systematic Review of Sources of Variability and Uncertainty in eDNA Data for Environmental Monitoring. Frontiers in Ecology and Evolution 8: 135. https://doi.org/10.3389/fevo.2020.00135
- Mauvisseau Q, Burian A, Gibson C, Brys R, Ramsey A, Sweet M (2019) Influence of accuracy, repeatability and detection probability in the reliability of species-specific eDNA based approaches. Scientific Reports 9(1): 580. https://doi.org/10.1038/s41598-018-37001-y
- McCarthy A, Rajabi H, McClenaghan B, Fahner NA, Porter E, Singer GAC, Hajibabaei M (2023) Comparative analysis of fish environmental DNA reveals higher sensitivity achieved through targeted sequence-based metabarcoding. Molecular Ecology Resources 23(3): 581–591. https://doi.org/10.1111/1755-0998.13732
- McElroy ME, Dressler TL, Titcomb GC, Wilson EA, Deiner K, Dudley TL, Eliason EJ, Evans NT, Gaines SD, Lafferty KD, Lamberti GA, Li Y, Lodge DM, Love MS, Mahon AR, Prefnder ME, Renshaw MA, Selkoe KA, Jerde CL (2020) Calibrating Environmental DNA Metabarcoding to Conventional Surveys for Measuring Fish Species Richness. Frontiers in Ecology and Evolution 8: 276. https://doi.org/10.3389/fevo.2020.00276
- McPhail JD (2007) The Freshwater Fishes of British Columbia. University of Alberta, Calgary Alberta, 620 pp. https://doi.org/10.1515/9780888648532
- Meng G, Li Y, Yang C, Liu S (2019) MitoZ: A toolkit for animal mitochondrial genome assembly, annotation and visualization. Nucleic Acids Research 47(11): e63. https://doi.org/10.1093/nar/gkz173
- Meusnier I, Singer GAC, Landry JF, Hickey DA, Hebert PDN, Hajibabaei M (2008) A universal DNA mini-barcode for biodiversity analysis. BMC Genomics 9(1): 214. https://doi.org/10.1186/1471-2164-9-214 [online]
- Minamoto T, Yamanaka H, Takahara T, Honjo MN, Kawabata Z (2012) Surveillance of fish species composition using environmental DNA. Limnology 13(2): 193–197. https://doi.org/10.1007/s10201-011-0362-4
- Muñoz-Pajares AJ (2021) Nosidier: Substitution and Indel Distances to Infer Evolutionary Relationships. https://rdrr.io/cran/sidier/
- O'Leary NA, Wright MW, Brister JR, Ciufo S, Haddad D, McVeigh R, Rajput B, Robbertse B, Smith-White B, Ako-Adjei D, Astashyn A, Badretdin A, Bao Y, Blinkova O, Brover B, Chetvernin V, Choi J, Cox E, Ermolaeva O, Farrell CM, Goldfarb T, Gupta T, Haft D, Hatcher E, Hlavina W, Joardar VS, Kodali VK, Li W, Maglott D, Masterson P, McGarvey KM, Murphy MR, O'Neill K, Pujar S, Rangwala SH, Rausch D, Riddick LD, Schoch C, Shkeda A, Storz SS, Sun H, Thibaud-Nissen F, Tolstoy I, Tully RE, Vatsan AR, Wallin C, Webb D, Wu W, Landrum MJ, Kimchi A, Tatusova T, DiCuccio M, Kitts P, Murphy TD, Pruitt KD (2016) Reference sequence (RefSeq) database at NCBI: Current status, taxonomic expansion, and functional annotation. Nucleic Acids Research 44(D1): D733–D745. https://doi.org/10.1093/nar/gkv1189
- Paradis E, Schliep K (2018) ape 5.0: An environment for modern phylogenetics and evolutionary analyses in R. Bioinformatics (Oxford, England) 35(3): 526–528. https://doi.org/10.1093/bioinformatics/bty633
- Piñol J, Mir G, Gomez-Polo P, Agustí N (2015) Universal and blocking primer mismatches limit the use of high-throughput DNA sequencing for the quantitative metabarcoding of arthropods. Molecular Ecology Resources 15(4): 819–830. https://doi.org/10.1111/1755-0998.12355
- R Core Team (2018) R: A Language and Environment for Statistical Computing.
- Ratnasingham S, Hebert PDN (2007) BOLD: The Barcode of Life Data System: Barcoding. Molecular Ecology Notes 7(3): 355–364. https://doi.org/10.1111/j.1471-8286.2007.01678.x
- Riaz T, Shehzad W, Viari A, Pompanon F, Taberlet P, Coissac E (2011) ecoPrimers: Inference of new DNA barcode markers from whole genome sequence analysis. Nucleic Acids Research 39(21): e145. https://doi.org/10.1093/nar/gkr732
- Ruppert KM, Kline RJ, Rahman MS (2019) Past, present, and future perspectives of environmental DNA (eDNA) metabarcoding: A systematic review in methods, monitoring, and applications of global eDNA. Global Ecology and Conservation 17: e00547. https://doi.org/10.1016/j.gecco.2019.e00547
- Ruskey JA, Taylor EB (2016) Morphological and genetic analysis of sympatric dace within the Rhinichthys cataractae species complex: A case of isolation lost. Biological Journal of the Linnean Society. Linnean Society of London 117(3): 547–563. https://doi.org/10.1111/bij.12657
- Sahu A, Kumar N, Singh CP, Singh M (2023) Environmental DNA (eDNA): Powerful technique for biodiversity conservation. Journal for Nature Conservation 71: 126325. https://doi.org/10.1016/j.jnc.2022.126325
- Sato Y, Miya M, Fukunaga T, Sado T, Iwasaki W (2018) MitoFish and MiFish Pipeline: A mitochondrial genome database of fish with an analysis pipeline for environmental DNA metabarcoding. Molecular Biology and Evolution 35(6): 1553–1555. https://doi.org/10.1093/molbev/msy074
- Schenekar T (2022) The current state of eDNA research in freshwater ecosystems: Are we shifting from the developmental phase to standard application in biomonitoring? Hydrobiologia 850: 1263–1282. https://doi.org/10.1007/s10750-022-04891-z
- Schroeter JC, Maloy AP, Rees CB, Bartron ML (2020) Fish mitochondrial genome sequencing: Expanding genetic resources to support species detection and biodiversity monitoring using environmental DNA. Conservation Genetics Resources 12(3): 433–446. https://doi.org/10.1007/s12686-019-01111-0
- Seeman T (2009) barnnap. https://github.com/tseemann/barrnap
- Shaw JLA, Clarke LJ, Wedderburn SD, Barnes TC, Weyrich LS, Cooper A (2016) Comparison of environmental DNA metabarcoding and conventional fish survey methods in a river system. Biological Conservation 197: 131–138. https://doi.org/10.1016/j.biocon.2016.03.010
- Simpson JT, Wong K, Jackman SD, Schein JE, Jones SJM, Birol İ (2009) ABySS: A parallel assembler for short read sequence data. Genome Research 19(6): 1117–1123. https://doi.org/10.1101/gr.089532.108
- Stein ED, Jerde CL, Allan EA, Sepulveda AJ, Abbott CL, Baerwald MR, Darling J, Goodwin KD, Meyer RS, Timmers MA, Thielen PM (2024) Critical considerations for communicating environmental DNA science. Environmental DNA 6(1): e472. https://doi.org/10.1002/edn3.472
- Taberlet P, Coissac E, Pompanon F, Brochmann C, Willerslev E (2012) Towards next-generation biodiversity assessment using DNA metabarcoding. Molecular Ecology 21(8): 2045–2050. https://doi.org/10.1111/j.1365-294X.2012.05470.x
- Taylor EB, Pollard S, Louie D (1999) Mitochondrial DNA variation in bull trout (Salvelinus confluentus) from northwestern North America: Implications for zoogeography and conservation. Molecular Ecology 8(7): 1155–1170. https://doi.org/10.1046/j.1365-294x.1999.00674.x
- Thomsen PF, Kielgast J, Iversen LL, Møller PR, Rasmussen M, Willerslev E (2012) Detection of a diverse marine fish fauna using environmental DNA from seawater samples. PLOS ONE 7(8): e41732. https://doi.org/10.1371/journal.pone.0041732
- Vences M, Lyra ML, Bina Perl RG, Bletz MC, Stankovic D, Lopes CM, Jarek M, Bhuju S, Geffers R, Haddad CFB, Steinfartz S (2016) Freshwater vertebrate metabarcoding on Illumina platforms using double-indexed primers of the mitochondrial 16S rRNA gene. Conservation Genetics Resources 8(3): 323–327. https://doi.org/10.1007/s12686-016-0550-y
- Wang S, Yan Z, Hanfling B, Zheng X, Wang P, Fan J, Li J (2021) Methodology of fish eDNA and its applications in ecology and environment. The Science of the Total Environment 755: 142622. https://doi.org/10.1016/j.scitotenv.2020.142622
- Wangensteen OS, Palacín C, Guardiola M, Turon X (2018) DNA metabarcoding of littoral hard-bottom communities: High diversity and database gaps revealed by two molecular markers. PeerJ 6: e4705. https://doi.org/10.7717/peerj.4705
- Westfall KM, Therriault TW, Abbott CL (2022) Targeted next-generation sequencing of environmental DNA improves detection of invasive European green crab (Carcinus maenas). Environmental DNA 4(2): 440–452. https://doi.org/10.1002/edn3.261
- Zaiko A, Pochon X, Garcia-Vazquez E, Olenin S, Wood SA (2018) Advantages and limitations of environmental DNA/RNA tools for marine biosecurity: Management and surveillance of non-indigenous species. Frontiers in Marine Science 5: 322. https://doi.org/10.3389/fmars.2018.00322
- Zaiko A, Greenfield P, Abbott C, von Ammon U, Bilewitch J, Bunce M, Cristescu ME, Chariton A, Dowle E, Geller J, Ardura Gutierrez A, Hajibabaei M, Haggard E, Inglis GJ, Lavery SD, Samuiloviene A, Simpson T, Stat M, Stephenson S, Sutherland J, Thakur V, Westfall K, Wood SA, Wright M, Zhang G, Pochon X (2022) Towards reproducible metabarcoding data: Lessons from an international cross-laboratory experiment. Molecular Ecology Resources 22(2): 519–538. https://doi.org/10.1111/1755-0998.13485
- Zinger L, Bonin A, Alsos IG, Bálint M, Bik H, Boyer F, Chariton AA, Creer S, Coissac E, Deagle BE, De Barba M, Dickie IA, Dumbrell AJ, Ficetola GF, Fierer N, Fumagalli L, Gilbert MTP, Jarman S, Jumpponen A, Kauserud H, Orlando L, Pansu J, Pawlowski J, Tedersoo L, Thomsen PF, Willerslev E, Taberlet P (2019) DNA metabarcoding—Need for robust experimental designs to draw sound ecological conclusions. Molecular Ecology 28(8): 1857–1862. https://doi.org/10.1111/mec.15060