title: Metagenome
design: Library preparation was performed at Genotypic Technologyﾒs Genomics facility. 25-100 ng of nanodrop quantified DNA was used for amplifying V3-V4 region of 16S region with specific primers which also have a ﾑtagﾒ sequence that are complementary to Illumina sequence adapter and index primers from the Nextera XT Index kit V2. This round of PCR generates single Amplicon of ~550 bp. The Amplified products were checked on the Agarose Gel before proceeding for Indexing PCR. In the next round of PCR (indexing PCR) Illumina sequencing adapters and dual Indexing barcodes are added using limited cycle PCR to give a final product of ~600 bp. The library was cleaned using HighPrep PCR (Magbio, Cat # AC-60050) magnetic beads and was Qubit quantified and validated for quality by running an aliquot on High Sensitivity Bioanalyzer Chip (Agilent).