title: 16S rRNA sequencing of microrganisms present in Fe(III)-reducing Ni-ISA degrading enrichment culture.
design: Sequencing of PCR amplicons of 16S rRNA was conducted with the Illumina MiSeq platform (Illumina, San Diego, CA, USA) targeting the V4 hyper variable regions (forward primer, 515F, 5'-GTGYCAGCMGCCGCGGTAA-3'; reverse primer, 806R, 5'-GGACTACHVGGGTWTCTAAT-3') for 2×150-bp paired-end sequencing. PCR amplification was performed using Roche FastStart High Fidelity PCR System in 50ul reactions under the following conditions: initial denaturation at 95°C for 2min, followed by 36 cycles of 95°C for 30s, 55°C for 30s, 72°C for 1 min, and a final extension step of 5min at 72°C. The PCR products were purified and normalised to 20ng each using the SequalPrep Normalization Kit . The PCR amplicons from all samples were pooled in equimolar ratios. The run was performed using a 4pM sample library spiked with 4pM PhiX to a final concentration of 10% following the method of Schloss and Kozich.
library_name: Iso-Ni 4