title: 16S rRNA gene amplicon sequences of bacterioplankton with Thermus thermophilus internal control: Amazon River plume water
design: DNA was extracted from bacterioplankton cells collected on filters following addition of Thermus thermophilus strain HB8 (ATCC) genomic DNA (0.01% by mass of the expected DNA recovery) to serve as an internal standard to account for variable extraction efficiency (Satinsky et al. 2013). Bacteria-specific PCR primers were used for amplification of the 16S rRNA V1-V2 regions: 27F with 454B FLX linker (GCCTTGCCAGCCCGCTCAG TC AGRGTTTGATYMTGGCTCAG) and 338R with 454A linker and unique 8 base pair barcode, denoted by N in primer sequence (GCCTCCCTCGCGCCATCAG NNNNNNN CA TGCWGCCWCCCGTAGGWGT).  Replicate amplicons were combined, quantified, pooled in normalized masses, purified, and pyrosequenced on a Roche-454 FLX Pyrosequencer at Engencore at the University of South Carolina using titanium chemistry (http://engencore.sc.edu).