title: 16SrRNA of wood-eating catfish guts
design: DNA was extracted from the contents of a 5 cm section of each frozen gut sample using a MoBio PowerSoil DNA extraction kit (MoBio, Carlsbad, CA). After an initial step in which the gut section was thawed (37 C) and vortexed for 2 min in 200 l of the first lysis buffer to disperse gut contents, DNA was extracted following the manufacturers specifications. Frozen woody debris samples were scrubbed with sterile toothbrushes into the first lysis buffer solution and DNA extracted from that suspension. DNA from all extractions was amplified using a dual index barcoding approach that targets the V4 regions of the 16S rRNA gene (Kozich, Wescott, Baxter, Highlander, & Schloss, 2013; Jackson, Stone, & Tyler, 2015). Amplification products were standardized using SequalPrep Normalization plates (Life Technologies, Grand Island, NY) and pooled prior to sequencing using an Illumina MiSeq platform at the Molecular and Genomics Core Facility at the University of Mississippi Medical Center.