Amplicon sequencing of Symbiodiniaceae ITS2: Amphisorus sp.
DNA was extracted using the DNeasy PowerSoil Pro Kit (QIAGEN). ITS2 region of the rRNA gene was amplified with Symbiodiniaceae-specific primers fused with Illumina adaptors: ITS-DINO 5-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-GTGAATTGCAGAACTCCGTG-3 [5] and ITS2Rev2 5-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-CCTCCGCTTACTTATATGCTT-3 [6]. PCR products were cleaned with AMPure XP beads (Beckman Coulter), and 5 L was used in the second PCR reaction with 2 M Nextera XT Index Primers and 2x KAPA HiFi HotStart ReadyMix. The second PCR products were cleaned with AMPure XP beads and sequenced on two lanes of Illumina MiSeq with 300 bp paired reads using a version 3 (600 cycle) reagent kit