16S rRNA 扩增子测序数据分析流程 - Illumina NovaSeq PE250 平台

The PE reads obtained from Illumina NovaSeq PE250 platform sequencing were first joined based on overlapping relationships. Simultaneously, fastp (v0.19.6) and FLASH (v1.2.7) were used to perform quality control and merging of the raw sequencing data, resulting in optimized data. DADA2 was then used to denoise the sequence, perform quality control, and filter the sequences to obtain amplicon sequence variants (ASV) representing sequence and abundance information. The silva138/16s_bacteria species annotation database was used as the reference sequence, and the ASVs were taxonomically annotated using the classifysklearn (Naive Bayes) algorithm in QIIME2, with a classification confidence set at 70%. The obtained sample information was analyzed using ASV analysis, which can be used for analyzing different indicators and further detecting sequencing depth. Based on taxonomic information, statistical analysis of community structure can be conducted at various taxonomic levels to obtain microbial community information for each sample.