16S rRNA amplicon sequencing of activated sludge microorganisms under exposure to triclosan

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 activated sludge microorganisms and quantify the relative abundance of activated sludge microorganisms under the exposure of triclosan. 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 three triclosan concentrations (0.1, 1 and 100 microgram/L)were selected to treat activated sludge microorganisms. The control group without triclosan was simultaneously set up. By comparing the relative abundance of bacteria (at both phylum and genus levels) from triclosan-treated groups and control group, the effects of triclosan on plasmid permissiveness to activated sludge microorganisms can be revealed.