n4_11330 (CRX1936861)
Genomic DNA was extracted from 5 ml of each saliva sample using the TIANamp Soil DNA Kit (DP336) according to the manufacturer's instructions. DNA concentration was quantified using a Qubit 2.0 fluorometer (Thermo Fisher Scientific, Waltham, MA, USA). The 16S rRNA gene was amplified using primers 341F (5'-CCTACGGGNGGCWGCAG-3') / 805R (5'-GACTACHVGGGTATCTAATCC-3'), targeting the V3-V4 hypervariable region. Sequencing was performed on an Illumina NovaSeq PE250 platform according to the manufacturer's protocol. Paired-end reads were demultiplexed using unique barcodes, and barcodes and primer sequences were removed. Reads were then merged using FLASH. Quality filtering of raw reads was performed with fqtrim (v0.94) to generate high-quality clean tags, and chimeric sequences were filtered using Vsearch software (v2.3.4). Amplicon sequence variants (ASVs) were inferred and dereplicated using the DADA2 algorithm, resulting in a feature table and representative feature sequences. Sequence alignment was performed using BLAST, and taxonomic annotation of ASVs was conducted using the SILVA database (release 138).  Alpha- and beta-diversity analyses were conducted using QIIME2.
NGDC
CRX1936861
Xiangya Hospital Central South University
DRX1006233