16SrRNA of stool among women in Punjab, India
DNA extraction was performed using QIAamp PowerFecal Pro DNA Kit and quantified using Qubit DNA High sensitivity Assay (Invitrogen), Qubit RNA Broad range Assay (Invitrogen) and QIAxpert. Samples were diluted to 5 ng and were amplified for the 16S region (1500bp) followed by the V3V4 region (460-480bp) using 16S and V3V4 primers respectively in a nested PCR approach. All PCR products were further processed for DNA library preparation using the Twist MF Library Prep Kit for Illumina. All prepared libraries were checked for fragment distribution using the 5300 Fragment Analyzer system. Prepared libraries were sequenced using the Illumina NextSeq2000 platform.