Transcriptomic response of Prymnesium parvum to glyphosate

Prymnesium parvum (UTEX-2797) was grown as batch culture on modified Artificial Seawater Medium (f/2, 5 psu), Photoperiod: 12 L: 12 D, Light intensity: 87 µmol s-1 m2, Temperature: 23.4 °C, for 9 days, which corresponds to a point near the end of the exponential growth phase, with technical-grade glyphosate at 0, 0.1, 2 or 4 mg/L. The entire volume of each replicate flask was centrifuged at 3600 rpm for 10 min at 8 °C. The supernatant was discarded by careful suctioning with a Pasteur pipette and the cell pellet was resuspended in Trizol reagentwith a vortex mixer. The suspension was preserved at -80 °C for later RNA extraction. Total RNA was extracted using the TRIzol®- Direct-zol™ RNA Kits according to the manufacturer's protocol, followed by DNAase I treatment. RNA quality was determined in Agilent TapeStation 2200 using RNA Screen Tape. The mRNA purification, RNA fragmentation, double stranded cDNA and adaptor ligation was generated using the TruSeq Stranded mRNA Library Prep #20020594. Libraries were spiked with 1% phiX libraries.