Microbiome analysis of cattle slurry amended soils using 16S amplicon sequencing
Genomic prokaryotic DNA of the V4 hypervariable region of the 16S rRNA gene was amplified using the barcoded primers described by Caporaso et al., 2011. These were 515f and 806r. All PCRs were performed in triplicate, using high fidelity Q5 taq polymerase (NEB) and pooled before being size selectively purified. Samples were quantified (Qubit) then pooled in equimolar concentrations before being sent to the Centre for Genomic Research (University of Liverpool) for paired-end sequencing Illumina MiSeq with 2 x 300bp V3 kit chemistry to a minimum depth of 100K