16S rRNA V4 region sequencing of human supragingival plaque
Total DNA extracted from supragingival plaque using PowerLyzer PowerSoil DNA Isolation Kits (Qiagen). PCR reactions held at 98C for 3 min, followed by 30 cycles of amplification at 98C for 10 s, 48C for 10 s, and 72C for 15 s. Barcoded amplicons were purified using Agencourt AMPure XP Reagent (Beckman Coulter), & quantified on Agilent 4200 TapeStation system (Integrated Sciences). Made 30 pM amplicon libraries, used IonChef System with the Ion 510 & Ion 520 & Ion 530 Chef Kit (Thermo Fisher Scientific) and Ion 520 Chip Kit (Thermo Fisher Scientific) with 200 bp chemistry.