16S amplicon sequencing of Anaxyrus americanus skin swabs: individual 52, ventral
We extracted DNA, combining duplicate swabs for single extractions, using a ZymoBIOMICS DNA/RNA Miniprep Kit according to the manufacturers protocol. We performed 16S rRNA illumina-tag PCR reactions on all 160 DNA extracts per the Earth Microbiome Projects protocol. We pooled PCR products in equimolar concentrations and gel-purified them on a 2% agarose gel using the QIAquick Gel Purification Kit. The purified pool was subsequently sequenced using Illumina MiSeq v2 kits to produce paired-end 250 base pair reads.