In silico PCR analysis identifying the completeness of the reference database and mismatches in the forward and reverse primer binding site for the fish (16S) and crustacean (16S) assay. Mismatches in the primer binding sites are indicated by coloured circles. * denotes the presence of a reference sequence without primer-binding regions, ** denotes newly-barcoded species and *** denotes species only detected by the eDNA survey. Species with missing primer information are a result of incomplete reference information in the database.