16S rRNA of Deer: feces
Targeted 16S rRNA amplicon sequencing was undertaken by Monash University Genomics and Bioinformatics Platform (Monash University) as previously described in Henry et al., (2016, 2018). Briefly, 16S rRNA PCR amplicon libraries were prepared by amplifying the V34 region of the bacterial rRNA gene in triplicate 50 L reactions. Reactions contained 1 m forward (TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG) and reverse (GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC) primers; 5 L of purified concentration-standardised genomic DNA; 25 L of 2x HiFi HotStart ReadyMix (Kapa Biosystems, USA). Amplified DNA, derived from the triplicate reactions, was then pooled for each sample and purified using Ampure XP (0.6 V) according to the manufacturer's protocol. Subsequently, the libraries were pooled in equimolar concentrations and sequenced using an MGITech DNBSEQ-G400RS sequencing instrument with High-Throughput Sequencing Set FCLPE100 chemistry using the manufacturer's instructions: MGI document SOP-013-B01-082 revision 7.