In silico evaluation of insect groups (after Misof et al. 2014) for selected metabarcoding primer pairs. COI reference sequences for primer evaluation were obtained from BOLD and NCBI using PrimerMiner and processed into OTUs (3% similarity). For primer-template mismatches, penalty scores were calculated using PrimerMiner (lower penalty score = better expected primer performance). The individual mean penalty scores are given in bar plots for each primer and insect order. The average penalty score was calculated for each primer for orders with at least 100 OTUs for the respective primer pair. The typically used primer combinations are indicated by connected grey lines on the left, as well as black lines for the newly developed primer pairs. Templates for primer development are indicated with blue boxes, while the newly developed primers are highlighted with blue backgrounds. The fwhR2 primer had a design error and is highlighted in grey.