GSM9179571: Sertraline hydrochloride exposure, replicate B, concentration 1960; Raphidocelis subcapitata; RNA-Seq
RNA was extracted from pooled homogenates (Plant/Fungi Total RNA Purification kits, #31900, Norgen Biotek, Ontario, Canada) with a protocol slightly modified from the manufacturer's instructions. First, the algae were centrifuged, and the supernatant was aspirated. Glass beads (0.5 mm) were added to each well and the plate went through five cycles of freezing with liquid nitrogen and bead milling for 2 minutes. Next, 400 µl of kit Buffer C was added to each well and then the manufacturer's procedure was followed. Libraries were prepared using the Lexogen QuantSeq 3' mRNA-Seq V2 Library Prep Kit with UDI (Unique Dual Index) and QuantSeq UMI (Unique Molecular Identifier) module with PhiX spike-in making up ~5% of the total reads in each lane. The UMI module (https://faqs.lexogen.com/faq/quantseq-umi-module) added a random, 6nt sequence to the start of each read 1 (R1), followed by a constant, 4nt (TATA) spacer. Preparation followed manufacturer's recommendations. Completed libraries were quality checked and quantified using Qubit dsDNA HS and Agilent 4200 TapeStation HS DNA1000 assays. Libraries were pooled in equimolar amounts and quantified using the Invitrogen Collibri Quantification qPCR kit.
GEO
GSM9179571_r1