Metabarcoding analysis of mammalian mitochondrial 16S genes: surface water
An index PCR was conducted to attach Nextera XT index primers to the amplicons. The final PCR products were then pooled and purified using Mag-Bind Total Pure NGS Kit (Omega Bio-Tek). The purified products were then gel electrophoresed, and the excised band was purified using the Promega Wizard SV Gel and PCR Clean-Up System (Promega). The double-purified PCR libraries were then analyzed by Agilent TapeStation for base pair size and a Qubit 3.0 Fluorometer for quantification. The resulting amplicon libraries were then used as templates for sequencing with the Illumina MiSeq System following the manufacturers instructions.