Microbiome of soil samples from different salamander habitats
Each sample's extracted total DNA was qunatified via Qubit and, when necessary, normalized prior to amplification. We then amplified full-length 16S rRNA gene regions using barcoded primers following the Amplification of Full-Length 16S Gene with Barcoded Primers for Multiplexed SMRTbell Library Preparation and Sequencing protocol for 27 cycles. Amplification products were visualized on a 1.2% agarose gel electrophoresis, pooled, and purified using AMPure PB beads prior to library construction. SMRTbell adapters were then ligated onto purified PCR products using SMRTbell Express Template Prep Kit 2.0. Libraries were sequenced on a PacBio Sequel IIe system at the University of Floridas Interdisciplinary Center for Biotechnology Research (ICBR).