eDNA of invertebrates
Primers targeting metazoans (mlCOIintF and jgHCO2198) were synthetized with locus-specific overhangs, and index specific primers were attached. PCRs and purification steps were executed. Final PCR products were measured with Qubit dsDNA HS Assay and average fragment size was estimated. Equimolar concentrations of PCR products were pooled together. Libraries were sequenced using an Illumina MiSeq v2 500-cycle Kit on an Illumina MiSeq platform.