GSM5550025: A41_A_11m (39-108 days post hatch dataset); Nothobranchius furzeri; RNA-Seq
RNA was extracted with modified Trizol protocol, using Qiazol lysis reagent (Qiagen). In brief, samples were lysed in 1mL QIAzol and 0.2mL of Chloroform was added. Samples were mixed and centrifuged at 20000g for 20 minutes at 4°C in a tabletop centrifuge. After the centrifugation, the upper aqueous phase was withdrawn and mixed with 1.1 volumes (550µl) of isopropyl alcohol, 0.16 volumes (80µl) of 2M, pH 4.2 Sodium acetate and 1µL (10µg) of GlycoBlue to precipitate RNA. Samples were then centrifuged at 20000g for 30 minutes at 4°C, washed twice with 80% ethanol and centrifuged at 7500g for 5 minutes at 4°C. The resulting pellet was dried for no more than 5 minutes and dissolved in nuclease-free water. Preparation of libraries was done using Illumina's TruSeq stranded mRNA kit according the manufacturer's description.