Fecal metabarcoding Anthus campestris
The eukaryotic mini-barcode miniB18S_81 designed by Cabodevilla et al. (2022) was used to amplify prey DNA, targeting the 18S rRNA gene. Negative PCR control was included in which no fecal DNA was added. A second PCR was performed to index each amplified product and attach Illumina adaptors. Resulting fragments were checked in Bioanalyzer before pooling in equimolar amounts. Library was further purified using AMPure XP beads (Beckman Coulter). The final library was sequenced in an Illumina MiSeq NGS platform with a MiSeq Reagent Kit v3-600 cycles, following the manufacturers instructions.