Shrew (Sorex sp.) Gut Contents Identified by DNA Metabarcoding, North America, 1968-2016

DNA from expertly identified helminths from fresh shrew gastrointestinal (GI) tracts was extracted using the Zymo Research (ZR) Genomic-DNA-tissue MicroPrep kit per manufacturer’s instructions (Zymo Research, Irvine, CA). PCR primers, conditions and Sanger sequencing reactions of these reference helminths used in Next Generation Sequencing (NGS) primer design are provided in Greiman et al (2018). Individual shrews with gastrointestinal (GI) tracts available for this study were identified using Arctos and obtained from the Museum of Southwestern Biology. DNA was extracted from GI tract contents using the ZR Fecal DNA MiniPrepTM kit with minor modifications outlined in Greiman et al (2018, 2020). Extractions were quantified with fluorometry and diluted to 50 ng/µl when possible for library preparation. NGS primers listed in Greiman et al (2018, 2020) were used to amplify targeted portions of helminths (16S mtDNA, 12S mtDNA, 28s rDNA) and bacteria (16S V4) in triplicate with positive and negative controls. Clean-up, library normalization, pooling and Illumina MiSeq parameters are all detailed in Greiman et al (2018, 2020).