Bacterial diversity of a terrestrial sulfidic spring outflow channel
DNA extraction was done in duplicate for each sample, utilizing the PowerSoil DNA Isolation Kit (MO BIO Laboratories, Carlsbad, California, USA), according to manufacturer instructions. DNA purity and concentration for each extraction were determined on a Gene_Quantll spectrophotometer (Amersdam Biosciences, Piscataway, NJ). Extractions were stored in TE buffer at -20 degrees C until use. DNA extractions were purified and sequenced by the Research and Testing Laboratories (RTL) (Lubbock, Texas, USA) using massively parallel bacterial tag-encoded FLX amplicon pyrosequences (bTEFAP), as described previously (Dowd et al., 2008a, 2008b) for the V1-V3 region of the 16S rRNA gene. Following sequencing, all failed sequence reads, low quality score sequences ( less than 20), and non-bacterial ribosomal sequences were removed.