Illumina 16S rRNA Sequencing of Bacterial Multispecies

Herein, a 10 mm × 1 mm glass disk was regarded as the blank group due to its insulative characteristic compared to the metal disks, excluding the potential electrochemical interferences. The universal reaction of V3-V4 regions of the bacterial 16S rRNA genes was amplified by 338F/806R primers. PCR was conducted according to the previous instructions (Cheng et al., 2019). The unprocessed data was obtained after the first sequencing with an Illumina Miseq sequencing platform (PE300, Personalbio). After the process of quality control, the resulting sequences were clustered into amplicon sequence variants (ASVs) at a 100% similarity using Quantitative Insights Into Microbial Ecology 2 (QIIME2) dada2 clustering. Diagrams were produced with various aspects based on the abundance of ASVs combined with a set of multivariate analyzing tools. A prediction of microbial metabolism functions was provided using PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of Unobserved States) tools according to the Kyoto Encyclopedia of Genes and Genomes (KEGG) database.

All 16S sequencing data on gene expressions have been deposited in NCBI’s Sequence Read Archive (SRA) and are accessible through accession number PRJNA707007¹.