GSM9574355: PFNA 10  ug/L, 1; Danio rerio; RNA-Seq
Total RNA was extracted using TRIzol (Thermo Fisher Scientific, Waltham, MA USA) and concentration was determined using the Qubit® 2.0 Fluorometer (Ther-moFisher, Grand Island, NY, USA). A total of 12 RNA samples were used for RNA-seq library construction (RIN > 7) (Agilent 2100 Bioanalyzer, Agilent Technologies, Inc.). Samples were collected following the 7-day exposure as outlined in Section 2.3. Each beaker of fish containing 14–15 individuals were pooled to generate a biological replicate. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using either dUTP for directional library. For the directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification