title: Root
design: 16S V3 Metagenome libraries were prepared using region-specific targeting proprietary primers. Genomic DNA was amplified for 21 cycles using KAPA HiFiHotStart PCR Kit. The forward and reverse primer concentration waskept at 5 uM each. The amplicons were analyzed on 1.2% agarose gel. 1ul of diluted round 1 PCR amplicons were used for Indexing PCR. Here, the round 1 PCR amplicons were amplified for 10 cycles to add Illumina sequencing barcoded adaptors. Round 2 PCR amplicons were analyzed on 1.2% agarose gel. Further the libraries were normalized and pooled for sequencing.