16S rRNA gene amplicon sequencing of the southern sea otter microbiota: PCR negative control
Sea otter samples were prepared for amplicon analysis as described in Bik et al., 2016. 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.