SSU rRNA gene amplicon sequencing V3-V4: pediatric subgingival plaque
Total bacterial genomic DNA was extracted from the oral biofilm samples using QIAamp DNA Mini kits (Qiagen) according to the manufacturer's instructions. PCR amplification of the hypervariable V3-V4 region of the 16S ribosomal RNA (rRNA) genes was performed using the universal bacterial primer pairs, 341F (5-ACTCCTACGGGAGGCAGCAG-3) and 806R (5-GGACTACHVGGGTWTCTAAT-3). PCR amplification and library construction were performed according to Illumina's standard protocol for 16S Metagenomic Sequencing Library Preparation (Part # 15044223 Rev. B). Paired-end MiSeq sequencing was performed on the amplicon libraries using an Illumina MiSeq system (300PE) at the Beijing Genomics Institute (BGI Institute, Shenzhen, China).