Metagenomic Sequencing of Hot Springs from Central India
The purified metagenomic DNA was used as a template for generating the 16S rRNA V3 amplicon library by optimised PCR amplification. The amplified product was evaluated on 2% w/v agarose gel and purified using Agencourt Ampure XP kit (Beckman Coulter, USA). Amplicon libraries were prepared by following the Illumina 16S metagenomic library preparation guide and evaluated on 2100 Bioanalyzer using Bioanalyzer DNA 1000 kit to estimate the library size and were quantified using Qubit dsDNA HS kit on a Qubit 2.0 fluorometer (Life technologies, USA). Equal concentration of libraries were loaded on Illumina NextSeq 500 platform using NextSeq 500/550 v2 sequencing reagent kit (Illumina Inc., USA) and 150X2 bp paired-end sequencing of the libraries was performed.