title: Microbial diversity in Puthuvype Mangrove Sediment, Cochin, India
design: 16S V3-V4 metagenome libraries were prepared using 16S Bacteria V3-V4 region-specific targeting proprietary primers at Genotypic Technology Pvt. Ltd., Bangalore, India. Briefly, 20 ng of genomic DNA (Quantification method: Nanodrop spectrophotometer) was amplified for 26 cycles using KAPA HiFi HotStart PCR Kit (KAPA Biosystems Inc., Boston, MA USA). The forward and reverse primer concentration is kept at 0.2 uM each. The amplicons were analyzed on 1.2% agarose gel. 1ul of round 1 PCR amplicons were used for Indexing PCR (round 2). Here, the round 1 PCR amplicons were amplified for 10 cycles to add Illumina sequencing barcoded adaptors (Nextera XT v2 Index Kit, Illumina, U.S.A.). Round 2 PCR amplicons were analyzed on 1.2% agarose gel (Figure 1: Round 2 PCR).