title: 16S rRNA seq of botteld water microbiota
design: DNA was extracted from bottled mineral water microbial communities collected with GTTP filters using a Phenol-Chloroform based method. From DNA samples the V4 to V6 region of the 16S rRNA gene was amplified with the primer pair 909F, 5'-ACTCAAAKGAATWGACGG- 3' / 1492R, 5'-NTACCTTGTTACGACT- 3'. These amplicons were used in as a template in a second PCR with a barcoded reverse primer (1492R). The amplicons were then purified with a AMPureXP solution and then quantified with PicoGreen DNA stain. Afterwards an equal amount of DNA from each sample was pooled and pyrosequencing of barcoded amplicon pools was performed with Titanium reagents on a 454 genome sequencer FLX.