metagenome of the sulfate reducing inoculum with Pure Glycerol as e-donor
The 16S rRNA gene V4 variable region PCR primers 515F (5'-GTGCCAGCMGCCGCGGTAA) and 806R (5'-GGACTACHVHHHTWTCTAAT) with barcode on the forward primer. Were used the HotStarTaq Plus Master Mix Kit (Qiagen, USA) under the established conditions: 94C for 3 minutes, followed by 30 cycles of 94C for 30 seconds, 53C for 40 seconds and 72C for 1 minute, after which a final elongation step at 72C for 5 minutes was performed. Was used 2% agarose gel to determine the success of amplification and the relative intensity of bands. The samples were pooled together in equal proportions based on their molecular weight and DNA concentrations and purified using calibrated Ampure XP beads. Then the pooled and purified PCR product was used to prepare Illumina DNA library. Sequencing was performed at MR DNA (www.mrdnalab.com, Shallowater, TX, USA) on a MiSeq following the manufacturers guidelines. Sequence data were processed using MR DNA analysis pipeline (MR DNA, Shallowater, TX, USA).
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