16S rRNA seq of soil metagenome-pre
DNA was extracted using the HiPurA Soil DNA Isolation Kit (Hi-Media, India) following the manufacturers protocol. High throughput 16S rRNA gene amplicon sequencing was performed. For this, DNA purity and concentration were confirmed using a NanoDrop 1000 spectrophotometer (Thermo Fisher Scientific, USA). Then the V3V4 hypervariable region of the 16S rRNA gene was amplified using V3_F (5'- CCTAYGGGRBGCASCAG -3') and V4_R (5'- GGACTACNNGGGTATCTAAT -3') primers. PCR Product quality was verified on the 2% agarose gel, and sequencing libraries were prepared using NEBNext Ultra II DNA Library Prep Kit for Illumina (Thermo Fisher Scientific, USA).