Evaluating Biofilm-Related Gene Expression

The effect of ZnO NPs on the biofilm-related genes (fnbA, fnbB, ebpS, icaC) [43 (link)] was determined by qRT-PCR, and the utilized housekeeping gene was 16S rRNA. The sequences of the primers are presented in Table S1. After extracting total RNA with the GeneJET RNA purification kit (Thermo Scientific, Waltham, MA, USA) described by the manufacturer, cDNA was formed using a power cDNA synthesis kit (iNtRON Biotechnology, Seongnam, Korea). Then, cDNA was amplified by Power SYBR^® Green master mix (Thermo Scientific, Waltham, MA, USA) in a Rotor-Gene Q 5plex machine (Qiagen, Hilden, Germany). We used the 2^−ΔΔCt method for the calculation of the relative gene expression. The expression of the genes in the isolates before treatment was 1 [44 (link)].