Fluid 16S
25 ng of DNA was used to amplify 16S rRNA hyper variable region V3-V4. The reaction includes KAPA HiFi HotStart Ready Mix and 100 nm final concentration of modified 341F and 785R primers. The PCR involved an initial denaturation of 95C for 5 min followed by 25 cycles of 95C for 30s, 55C for 45s and 72C for 30s and a final extension at 72C for 7 min. The amplicons were purified using Ampure beads to remove unused primers. Additional 8 cycles of PCR was performed using Illumina barcoded adapters to prepare the sequencing libraries.