Environmental DNA (eDNA) COI metabarcoding Illumina MiSeq NGS PCR Protocol v1

This protocol is aimed at amplifying the cytochrome c oxidase subunit I (COI) mitochondrial gene in eukaryotes. The primers (forward: mlCOIintF, reverse: HCO2198) utilized in this protocol are based on the primers utilized in Leray et al. 2013 (forward) and Folmer et al. 1994 (reverse). Amplicons generated using this protocol can then be sequenced using the Illumina platform. Primers used: Fluidigm CS1+mlCOIinfF Fluidigm CS2+HCO2198 Secondary COI PCR primers PE1-BC-CS1 PE2-BC-CS2