Barcode reference library coverage analyses

In silico primer evaluation

Sampling, identification and laboratory processing of specimens

Mock community design

Metabarcoding PCR, replication strategy, library preparation and sequencing

Quality filtering, MOTU clustering and taxonomic assignment

Barcode coverage

Species delimitation congruence

In silico primer evaluation

Bioinformatic analysis of mock communities and detection rates

Barcode coverage analysis

Congruence of MOTUs with morphospecies

Pipeline evaluation and potential error sources

Quantitative estimations from metabarcoding results: Is it possible?

Multiple MOTUs originating from single specimens