title: 16S amplicon sequencing of metagenome: Red Sea marine sediment
design: DNA was extracted using the PowerSoil Pro Kit (Qiagen). DNA was amplified targeting the V3-V4 region of the 16S SSU rRNA gene using the 341F/805R primer set (Herlemann et al., 2011) with the following PCR thermal protocol: (1) an initial denaturation step at 98C for 2 minutes, followed by 28 cycles of 98C for 20 s, 54C for 20 s, and 72C for 15 s, with a final elongation step at 72C for 2 min. PCRs were run in triplicates to reduce PCR-related bias, and successful amplification were verified via gel electrophoresis. The three technical replicates were then pooled and cleaned using AMPure XP Beads (Beckman Coulter). Sequencing libraries were prepared using the Illumina Nextera XT kit and cleaned using AMPure XP Beads.