16S sequencing of  Hesperophylax sp.: gut microbiome
We used the Zymo Quick-DNA Fecal/Soil Microbe 96 Kit for DNA extraction. Libraries were prepared as 20 L PCR reactions using dual-barcoded primers. We used the Just-a-Plate 96 PCR Normalization and Purification Kit for PCR product normalization. Samples were pooled in equal volumes and concentrated to 12 L using the gDNA Clean & Concentrator -5 Kit. We selected DNA fragments by size using a Select-a-Size DNA Clean & Concentrator Kit. Fragment sizes were evaluated using an Agilent Bioanalyzer, and fragment size selection was performed using a Sage Science Blue Pippin. The library was diluted to 4.0 pM and pooled with 15% PhiX. 600 uL of this solution was loaded into the MiSeq Reagent Kit v2 500 cycle cartridge. The V4 region of 16S rRNA was sequenced.