Efficient and stable metabarcoding sequencing data using DNBSEQ-G400 sequencer validated by comprehensive community analyses

Metabarcoding has become a widely used method for fast characterizing microbial communities in complex environmental samples. Here we demonstrate a high-throughput metabarcoding approach on microbial community assignment using DNBSEQ-G400 sequencing developed by MGI Tech. The accuracy of DNBSEQ-G400 on quantifying relative abundance was evaluated by bacterial and fungal mock samples, as well as using fungal mocks with various mixing ratios. Sequencing throughput and consistency were characterized by sequencing the fungal ITS2 region from 1144 soil samples as well as technical replicates of three randomly picked samples. DNBSEQ-G400 showed high accuracy in reproducing mock communities containing different proportions of bacteria and fungi. The taxonomic profiles of the 1144 soil samples generated by two DNBSEQ-G400 sequencing modes closely resembled each other. Sequencing across two platforms, DNBSEQ-G400 and MiSeq also showed similar patterns of beta diversity. These datasets provided to allow further study of utility, capacity and bias in short-read metabarcoding of microbial communities.