Illumina MiSeq paired end sequencing of SAMD00636938
DNA extraction was performed using QIAamp PowerFecal Pro DNA Kits (QIAGEN, Hilden, Germany). The extracted DNA samples were used as DNA templates in a polymerase chain reaction (PCR) performed in a reaction mixture of template DNA, KAPA HiFi HotStart Ready Mix, and primers specific for the 16S rRNA V3?V4 region under the following conditions: initial denaturation at 95?C for 3 min, followed by 35 cycles at 95? for 30 s, 55? for 30 s, and 72? for 30 s. The final elongation step was performed at 72? for 5 min. The amplicons were purified using AMPure XP beads (Beckman Coulter, Brea, USA) and attached to dual indices by index PCR using the Nextera XT Index kit (Illumina). Libraries were purified using AMPure XP beads (Beckman Coulter). The purified libraries were diluted to 4 nM in Tris?HCl buffer and pooled.