16S rRNA gene amplicon sequences of bacteria from soil leachate experiment
DNA was extracted from bacterioplankton cells collected on filters. Bacteria-specific PCR primers were used to amplifiy the 16S rRNA gene V4 region.  Primer design: (Illumina linker)-(barcode)-(pad)-(linker)-(16S primer).  The two primers used were 515f (AATGATACGGCGACCACCGAGATCTACAC-NNNNNNNN-TATGGTAATT-GT-GTGCCAGCMGCCGCGGTAA) and 806r (CAAGCAGAAGACGGCATACGAGAT-NNNNNNNN-AGTCAGTCAG-CC-GGACTACHVGGGTWTCTAAT).  Amplicons were purified and pooled in normalized masses using SequalPrep plates (Qiagen), and sequenced on an Illumina MiSeq sequencer at the Center for Genome Research and Biocomputing at Oregon State University (http://cgrb.oregonstate.edu).  Unique pairs of 8 bp barcodes associated with PCR primers were used for each sample, and these barcodes were used to demultiplex the output data from MiSeq sequencing.
SUB3102512
DNA2013-259