COI metabarcoding of Doryteuthis sanpaulensis
PCR was carried out in 25 l volumes, with 1X Q5 High-Fidelity master mix, 1X Q5 enhancer and 10 M of each primer. The annealing temperature was 45 C. Amplicons were visualized on agarose gels and quantified using the ImageLab Software v6.0. Based on the quantification, uniform amounts of each amplicon were merged into new tubes using a Biomek 4000 liquid handling robot. The DNA library was cleaned using 1.0X AMpure beads, and prepared using the NEBNext Fast DNA Library Prep Set for Ion Torrent (NEB). The amplified libraries were size-selected using BluePippin. The final libraries were quantified on a Fragment Analyzer instrument using the High Sensitivity Genomic DNA Kit. The libraries were sequenced on two 530 chips on an Ion GeneStudio S5 system.