Control7
After carrying out quantity and quality checks, 16S rRNA sequencing was carried out. The sequencing library was prepared primers which results in an amplicon whose size is around 400-450 bp. Library construction involved random fragmentation of the DNA sample combined with 5' and 3' adapter ligation. The adapter-ligated fragments were then PCR amplified and gel purified. The DNA samples were amplified using the universal primers that target the 16S region (v3-v4): Bakt_341F: CCTACGGGNGGCWGCAG and Bakt_805R: GACTACHVGGGTATCTAATCC. For cluster generation, the library was loaded into a flow cell where the generated fragments were acquired on a lawn of surface-bound oligos complementary to the library adapters. This resulted in the generation of distinct, clonal clusters through bridge amplification. When cluster generation was complete, the templates were sequenced using the Herculase II Fusion DNA Polymerase Nextera XT Index Kit V2 (Aligent, CA, United States) and the Next Generation Sequencing (NGS) Illumina MiSeq platform.