GSM4368859
GSM4368859: RNAseq_Syn_WH7803_LL_LT_J0-5_C1; Synechococcus sp. WH 7803; RNA-Seq
RNA extractions were performed from 150 mL of culture harvested by centrifugation for 7 min at 10,414 × g and 4°C in an Eppendorf 5804R, followed by 2 min centrifugation at 20,817 × g in Eppendorf tubes and 4°C in an Eppendorff 5417R centrifuge. Cell pellet was then resuspended in 1 mL Qiazol (Qiagen, Valencia, CA, USA) and quickly frozen in liquid nitrogen. After thawing the tubes at 65 °C, followed by and additional incubation of 5 min at this temperature, RNA was extracted using the Direct-Zol kit (Zymo Research Corp, Proteigene) as recommended by the manufacturer. Three successive DNase treatments were performed on the Zymo-Spin™-IIC Column using the Qiagen RNase-free DNase Set (Qiagen), followed by elution from the column in 30 L DEPC-treated water. RNA quantification and quality check were performed using a Qubit (Invitrogen) and a BioAnalyzer 2100 with the RNA 6000 Nano Kit (Agilent, Santa Clara, CA, USA), respectively. RNA samples were then frozen in liquid nitrogen and stored at -80°C. Ribodepletion was performed at the Genotoul platform (Toulouse, France) from 5 µg of RNA using the Ribo-Zero Bacteria Magnetic Kits (Illumina) in presence of RiboGuard RNase Inhibitor (Epicentre) and treated RNA samples were purified and concentrated using RNA Clean & Concentrator™-5 columns (Zymo research). 0.5 µL of ribo-depleted RNA was then used for quantification (Qubit RNA HS Assay Kit) and quality check (Bioanalyzer RNA Pico kit or Labchip GX). Libraries were then prepared using 100 to 400 ng of RNA with the Illumina TruSeq Stranded mRNA kit and sequenced as 150 bp paired-end reads on Illumina HiSeq 3000 System (Illumina, San Diego, CA, USA) at Genotoul (Toulouse, France). 18.5 +/- 4.7·10^6 paired-end reads were generated per sample.
GEO
GSM4368859
GEO Accession
GSM4368859