16S rRNA amplicon sequencing of mice faecal bacteria under exposure to artificial sweeteners

The goals of this study are to use MiSeq Sequencing System (Illumina, V3 300bp paired-end reads) to identify the spread of plasmid-mediated antibiotic resistance among gut microbiota and quantify the relative abundance of gut bacteria under the exposure of artificial sweeteners (saccharin, sucralose, aspartame, and acesulfame potassium). The V3-V4 regions were targeted and amplificated using the 341F-806R primers. The PCR amplicons were purified using Agencourt AMPure XP beads (Beckman Coulter). The 16S rRNA library was constructed by following Illumina workflow (# 15044223 Rev. B). A 300 mg/L) of four artificial sweeteners was selected to treat faecal bacteria. The control group without artificial sweeteners was simultaneously set up. By comparing the relative abundance of bacteria (at both phylum and genus levels) from artificial sweetener-treated groups and control group, the effects of artificial sweeteners on plasmid permissiveness to faecal bacteria can be revealed.