title: Metabarcoding of moose and deer feces: Folmer primers
design: Using the QIAamp PowerFecal Pro kit, we extracted DNA  following the manufacturers protocol. We quantified the DNA extract concentration with the Qubit dsDNA Broad Range Assay Kit. After DNA extraction, we submitted samples to the University of Minnesota Genomics Center. UMGC cleaned the extracted DNA with the AxyPrep Mag PCR Clean-Up Kit to remove PCR inhibitors. Utilizing Folmer primers (LCO1490: 5'-ggtcaacaaatcataaagatattgg-3' HC02198: 5'-taaacttcagggtgaccaaaaaatca-3, Folmer, Black, Hoeh, Lutz, & Vrijenhoek, 1994), UMGC performed PCR the extracted DNA. UMGC cleaned the PCR product with AMPure XP beads at a concentration of 1.8X the sample. UMGC then assessed the success of the PCR reaction with Agilent Tapestation and only proceeded with library preparation on samples that had bands from 600-1,200 bp. During library preparation, UMGC added index sequences and Nextera adapter sequences to the PCR product following standard protocols. They then used AMPure XP beads at a concentration of 0.65X to size select for fragments >300 bp. UMGC then sequenced the samples with the Illumina MiSeq with a paired-end 300 bp run.