Illumina NovaSeq 6000 paired end sequencing; Transcriptome responses in copepods Calanus finmarchicus exposed to phenanthrene and benzo[a]pyrene
Transcriptome responses in copepods Calanus finmarchicus exposed to phenanthrene and benzo[a]pyrene
Copepods were collected during sampling cruise in the northern Barents Sea in August 2019 and exposure was performed onboard. The copepod species were captured with a Bongo Net (180 μm mesh size), or a WP-3 net (1000 μm mesh size).The copepods were quickly transferred to a bucket with seawater and kept well diluted at 0−4°C until sorting to species. Calanus finmarchicus copepodite stages CV were hand-picked for exposure. For exposure a sample of 20 individuals was kept in a 0.5 L glass flask filled with 0.4 L filtered seawater and exposure was performed for 72 hours at a temperature of 3.5 ˚C to solvent control (DMSO), 0.1 µM BaP or 0.1 µM Phe in filtered seawater (0.7 µm). For each treatment group, 4-5 replicate flasks (n = 5) were used. After 72 hours, the animals were harvested and snap-frozen in liquid nitrogen for storage at -80 °C until RNA-extraction. Total RNA was isolated from frozen slices using TRI Reagent according to the manufacturer's protocol (Sigma-Aldrich, St. Louis, MO, USA). RNA concentration and quality were assessed using NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA) and Agilent 2100 bioanalyzer (Agilent Technologies, Santa Clara, CA, USA).  Performed by Novogene on Illumina NovaSeq 6000 system