Microbial diversity analysis of environmental samples

To analyze microbial diversity of environmental samples, the total DNA is extracted using in-house protocol or commercial kits, followed by DNA quantification via Qubit4.0 fluorometer. A variable region of 16S rRNA gene (e.g. V4) is amplified by Polymerase Chain Reaction (PCR) with barcoded primers. Then the purified V4 amplicon is subjected to next-generation sequencing technologies. The downstream bioinformatic analyses is achieved using DADA2 software for generation of amplicon sequence variant (ASV) and phyloseq package in R for data visualization.