OCUBMT045_day21
The 16S rRNA V3V4 region was PCR-amplified using specific primers over 20 cycles, and the PCR products were purified with Agencourt AMpure beads (Beckman Coulter, Tokyo, Japan). Then, the overhang and index sequences required for sequencing were added by a second-round PCR using NEBNext multiplex Oligos for Illumina (Dual Index Primers Set1, New England Biolabs, Ipswich, MA, USA) over eight cycles, and the products were purified with Agencourt AMpure beads.