16S rRNA sequencing of Submerged Low N
DNA was extracted from the samples using the DNeasy Powersoil Pro Kit (QIAGEN, India) according to the manufacturers protocol. Quantitative & qualitative analysis was done by the fluoroscent (Qubit fluorometer4.0) & agarose gel electrophoresis. The amplicons of V3-V4 were prepared using the forward primer (16S V3-V4 forward: CCTACGGGNGGCWGCAG) and the reverse primer (16S V3-V4 Reverse: GACTACHVGGGTATCTAATCC) with a KAPA HiFi HotStart Ready Mix (Roche, Basel, CHE). The sequencing libraries were prepared according to the  NEBNext Ultra II DNA Library Prep Kit for Illumina (New England Biolabs, USA). Dual index adapters for sequencing on the Illumina MiSeq platform were attached using the NEBNext Multiplex Oligos for Illumina(New England Biolabs, USA). These libraries were sequenced in a 250bp paired-end run for V3-V4 using the MiSeq Reagent Kit v3 (250 cycles).