Soil sample 2
The amplicon libraries were prepared using Nextera XT Index Kit (Illumina inc.) as per the 16S Metagenomic Sequencing Library preparation protocol (Part # 15044223 Rev. B). Primers for the amplification of the bacterial specific region were designed and synthesized at Eurofins Genomics Lab (16S rRNA forward primer 5-GCCTACGGGNGGCWGCAG-3 and 16S rRNA reverse primer 5-ACTACHVGGGTATCTAATCC-3). Amplification of the bacterial 16S region was carried out using PCR. 3l of PCR product was resolved on 1.2% Agarose gel at 120V for approximately 60 min or till the samples reached 3/4th of the gel. The QC passed amplicons with the Illumina adaptor were amplified using i5 and i7 primers that add multiplexing index sequences as well as common adapters required for cluster generation (P5 and P7) as per the standard Illumina protocol. The amplicon libraries were purified by AMPure XP beads and quantified using Qubit Fluorometer.