Data from: The choice of universal primers and the characteristics of the species mixture determines when DNA metabarcoding can be quantitative.
Pinol, Josep; Senar, Miquel Angel; Symondson, William O. C. (2018), Data from: The choice of universal primers and the characteristics of the species mixture determines when DNA metabarcoding can be quantitative., Dryad, Dataset, https://doi.org/10.5061/dryad.q2r3b1f
DNA metabarcoding is a technique used to survey biodiversity in many ecological settings, but there are doubts about whether it can provide quantitative results, i.e. 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.
Basically, 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. 1200 insect species.
The results showed that some primers pairs produced quantitative results under most circumstances, whereas some other primers failed to do so. Many species, and a high diversity within the mixture, helped the metabarcoding to be quantitative. 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.