South Suaeda 2021
The 16S rRNA gene PCR primers 27F/1492R  with barcode on the forward primer were used in a 35-cycle PCR  (5 cycle used on PCR products) using the HotStarTaq Plus Master Mix Kit (Qiagen, USA) under the following conditions: 94C for 3 minutes, followed by 35 cycles of 94C for 30 seconds, 53C for 40 seconds and 72C for 90 seconds, after which a final elongation step at 72C for 5 minutes was performed.  After amplification, PCR products are checked in 2% agarose gel to determine the success of amplification and the relative intensity of bands. Multiple samples are pooled together (e.g., 100 samples) in equal proportions based on their molecular weight and DNA concentrations. The PCR pool is then purified using Ampure PB beads (Pacific Biosciences).