Metatranscriptomic sequencing of environmental RNA from zebrafish water exposed to chlorpyrifos and multiple concentrations of carbendazim.

Adult zebrafish (Danio rerio) were exposed to carbendazim, chlorpyrifos, or their mixture at environmentally relevant concentrations for 96 hours in static tanks. Each tank held ten fish (five males and five females) without feeding or aeration. After exposure, 500 mL of water was collected from each tank and filtered through sterile 0.45 micron membranes. Filters were placed into cryovials, frozen in liquid nitrogen, and stored at -80 C. RNA on the filters was stabilized using DNA/RNA Shield and processed with ZymoBIOMICS BashingBead lysis tubes. Total RNA was extracted using the ZymoBIOMICS DNA/RNA Miniprep Kit. RNA samples were submitted to Guangdong Magigene Biotechnology Co., Ltd. for library preparation and sequencing. Sequencing was performed on the Illumina NovaSeq 6000 platform using paired-end 150 bp reads.