GSM2943819
GSM2943819: Control condition 3; Tetraselmis suecica; RNA-Seq
Cells were lysed using half a spatula of glass beads (about 200 mg; Sigma-Aldrich, Milan, Italy) for each 2 mL tube, incubating and mixing tubes for 10 min at 60°C and maximum speed in the Thermo Shaker BS100 (Biosan). RNA was then extracted using the Direct-zolTM RNA MiniPrep (Zymo Research), following the manufacturer's instructions. RNA quantity and purity were assessed by Nano-Drop (ND-1000 UV-Vis spectrophotometer; NanoDrop Technologies) monitoring the absorbance at 260 nm and the 260/280 nm and 260/230 nm ratios (both ratios were about 2.0). RNA quality was evaluated by gel electrophoresis that showed intact RNA, with sharp ribosomal bands. Total RNA quality was evaluated by measuring the RNA Integrity Number (RIN) with Agilent 2100 Bioanalyzer (Agilent Technologies, Inc.); high quality (RIN >8) RNA was used for the RNAseq. RNA-Seq libraries were constructed from 2.5 µg of total RNA using the Illumina TruSeq® Stranded mRNA kit (Illumina Inc., San Diego, CA, USA) according to manufacturer instructions. The libraries were size selected using the Pippin Prep automated gel electrophoresis system (Sage Science, Beverly, MA, USA) for 350 to 550 bp. Paired-end sequencing (100 bp × 2) was performed with the HiSeq 1000 Illumina platform using the TruSeq SBS v3-HS kit (200 cycles) and TruSeq PE Cluster v3-cBot-HS kit (Illumina).
GEO
GSM2943819
GEO Accession
GSM2943819