16S amplicon libraries from Rana sierrae skin swabs and water samples
Skin-surface associated microbes were collected using sterile synthetic swabs. Planktonic bacteria present in water were collected by 0.22 micron filtration of water samples. The V1-V2 regions of the 16S ribosomal RNA gene were amplified using primers 8f (AGRGTTYGATYMTGGCTCAG) and 338r (TGCWGCCWCCCGTAGGWGT) with Roche FLX Amplicon Adapters. Samples were multiplexed using 8-bp barcodes. PCR products were purified, pooled in equimolar quantities, and sequenced on a Roche/454 GS FLX using Titanium Chemistry.