title: DNA Metabarcoding ichthyoplankton
design: Total genomic DNA was obtained by Green and Sambrook protocol; Amplification of COI using different sets of primers in a two-step Polymerase Chain Reaction (PCR). Half of primers were Ward et al. (2005) and Jennings et al. (2019), the others were designed specifically for the present study; all contain adapters for the P5 and P7 regions for Illumina sequencing platform; Internal primers contain degenerate bases, to enhance capacity of identify a broad range of species. Two PCR steps involve the amplification of the COI gene in fragments of approximately 450 bp. This approach aims to optimize the sequencing by using 2 x 250 bp or 2 x 300 bp cartridges (Illumina Miseq), which ensures a minimum overlap of 30 bp between the pairs of reads and provides complete coverage of the target fragment.