COI metabarcoding of freshwater benthos: AD amplicon
Ethanol was removed by evaporation, homogenized in a blender, subsampled, extracted using a NucleoSpin tissue kit following a modified Machery-Nagel protocol, first round of PCR used untailed primers, second round used Illumina-tailed primers, purified amplicons were dual indexed for Illumina MiSeq sequencing