The choice of universal primers and the characteristics of the species mixture determine when DNA metabarcoding can be quantitative
Piñol, Josep (Centre de Recerca Ecològica i d'Aplicacions Forestals)
Senar Rosell, Miquel (Universitat Autònoma de Barcelona)
Symondson, William O.C. (Cardiff University)

Fecha:	2019
Descripción:	13 pàg.
Resumen:	DNA metabarcoding is a technique used to survey biodiversity in many ecological settings, but there are doubts about whether it can provide quantitative results, that is, the proportions of each species in the mixture as opposed to a species list. While there are several experimental studies that report quantitative metabarcoding results, there are a similar number that fail to do so. Here, we provide the rationale to understand under what circumstances the technique can be quantitative. In essence, we simulate a mixture of DNA of S species with a defined initial abundance distribution. In the simulated PCR, each species increases its concentration following a certain amplification efficiency. The final DNA concentration will reflect the initial one when the efficiency is similar for all species; otherwise, the initial and final DNA concentrations would be poorly related. Although there are many known factors that modulate amplification efficiency, we focused on the number of primer-template mismatches, arguably the most important one. We used 15 common primers pairs targeting the mitochondrial COI region and the mitogenomes of ca. 1,200 insect species. The results showed that some primers pairs produced quantitative results under most circumstances, whereas some other primers failed to do so. In conclusion, depending on the primer pair used in the PCR amplification and on the characteristics of the mixture analysed (i. e. , high species richness, low evenness), DNA metabarcoding can provide a quantitative estimate of the relative abundances of different species.
Ayudas:	Ministerio de Economía y Competitividad TIN2014-53234-C2-1-R Agencia Estatal de Investigación TIN2017-84553-C2-1-R
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Lengua:	Anglès
Documento:	Article ; recerca ; Versió acceptada per publicar
Materia:	COI ; Diet analysis ; Environmental DNA ; In silico PCR ; Insects ; Primer bias
Publicado en:	Molecular ecology, Vol. 28, Num. 2 (January 2019) , p. 407-419, ISSN 1365-294X

DOI: 10.1111/mec.14776
PMID: 29939447

Postprint 35 p, 1.1 MB

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Documentos de investigación > Documentos de los grupos de investigación de la UAB > Centros y grupos de investigación (producción científica) > Ciencias > CREAF (Centre de Recerca Ecològica i d'Aplicacions Forestals)
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