Miseq of Site D
Qiagen Power Soil Pro kit quipped with Inhibitor Removal Technology (IRT) was used for metagenomics DNA isolation. The V3-V4 (Product size ~459bp) region of 16s RNA gene was amplified using specific primers. The quantity and quality check (QC) of the amplified libraries were analyzed on TapeStation 4150 (Agilent Technologies) using High Sensitivity D1000 ScreenTape as per manufacturer instructions. Libraries were quantified using Qubit HS and qPCR mix pooled together for sequencing. The pooled PCR products (library) were loaded on sequencer for cluster generation by hybridization onto the oligonucleotide-coated surface of the flowcell. Immobilized DNA template copies were amplified by bridge amplification to generate clonal DNA clusters. Sequencing was performed usingPE300 kit for sequencing of 16S sequencing on Miseq platform. Raw reads obtained through Miseq platform were further processed in QIIME2 (Quantitative Insights into Microbial Ecology) pipeline (ver. 2020.11). to get good quality reads (QC>30) and Amplicon sequence variants(ASVs) the denoising and quality filtering was done using q2-dada2 plugin of QIIME2 pipeline.  The final reads were classified using QIIME feature-classifer command for taxonomy assignment