16S rRNA gene amplicon sequencing of the southern sea otter microbiota: fecal sample
Genomic DNA was extracted using the QiaAMP DNA Mini Kit (Qiagen, Valencia, CA, Cat No. 51304) as described in Bik et al. (2016). In-depth details on PCR amplification and library preparation can be found in the Supplemental Methods. Briefly, PCR amplification was performed in triplicate using 515F and 806rB primers with the 5 Prime Hot Master Mix (Quantabio, MA, Cat No. 2200410). Amplicons were sequenced with two 2 x 250 nt Illumina HiSeq 2500 lanes at the W.M. Keck Center for Comparative Functional Genomics at the University of Illinois, Urbana-Champaign (USA). One lane of sequencing was generated for the analysis of wild sea otters in this study, and a second for all samples used in other amplicon-based analyses.