Primers used for this study.

COI+ includes the “Folmer reigon” of COI along with the intergenic spacer between COI and COII, which is NCR2 of Smith et al.[7] and is coded as igr1 in McFadden et al.[24]. All PCRs began with a 2-min hot start at 94–96°C followed by the denaturing and annealing protocols below. Each cycle was completed with a 1 min at 72°C step (45 s for OctoH/L primers) for the number of cycles listed in the Table and after all cycles, a single round of 72°C for 5 mins followed by a 4°C hold. Where conditions are variable, conditions used most often are in bold, with adjustments to time and temps to get stragglers also shown.