Approach example pros cons
Combining many metabarcodes in the same study
Li et al. (2023) Analyzed freshwater biodiversity using four primers, focusing on bacteria; micro-eukaryotes; insects and fish Good coverage of biodiversity Resolution can be high for the selected taxa Costly Some taxon will always be missing
Universal markers
Holman et al. (2021) performed a joint biogeographical analysis of marine animals, protists and bacteria
Relatively cheap In principle, might cover the whole tree of life
Amplification rate and resolution are often heterogeneous across taxa
Combining universal and specific metabarcodes
Bloor et al. (2021) combined three universal (bacteria, eukaryotes, fungi) and four specific (seed plants, insects, springtails and earthworms) markers for a multi-trophic analysis of soil diversity
Good information on key groups Reduces the number of unrepresented taxa Costly Resolution can be strongly heterogeneous across taxa
Multiplex of primers
Govender et al. (2022) used six primer cocktails to analyze the diversity of 14 zooplankton taxa
Potentially excellent resolution Potentially excellent coverage of the tree of life Cheaper than analyzing each taxon separately Methodological developments required to optimize the multiplex Bioinformatics challenges
Shotgun sequencing
Pedersen et al. (2016) used ancient DNA to reconstruct post-glacial colonization patterns of plants, mammals and fish
Bypasses many limitations of metabarcoding (amplification, abundance) Can exploit the whole genomic DNA Can cover the whole tree of life Allows authentication of ancient eDNA Assignation heavily depends on reference databases Very costly Complex analytical pipelines