16S rRNA gene amplicon sequencing of dolphin oral swab sample
Genomic DNA was extracted from dolphin oral samples and 32 negative (PBS) controls using the DNeasy UltraClean 96 Microbial Kit (Qiagen Cat. #10196-4) following manufacturers instructions. The 16S V4 region was amplified using 515F and 806rB primers using Platinum II HotStart PCR Master Mix (ThermoFisher Cat. #14000013). The PCR products were pooled at equal volume and gel-purified. Final purification was performed using Macherey-Nagel NucleoSpin Gel and PCR Clean-up, Mini Kit (Fisher, Cat. #740609). Amplicons were sequenced on the Illumina MiSeq platform with 250-bp paired reads at the Stanford Chan Zuckerberg Biohub Facility.