Bacterial diversity along a transect of a sulfidic spring
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°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 (<20), and non-bacterial ribosomal sequences were removed.