GSM8679663: Intestine, no diet, 10 weeks, rep1; Nothobranchius furzeri; RNA-Seq
Total RNA from snap-frozen tissues was extracted in TRI-Reagent (Sigma) using TissueLyser LT (Qiagen). Phase separation was performed in Phasemaker™ Tubes (Invitrogen) and RNA purification, including a DNase I digestion step, was accomplished using the ReliaPrep™ RNA Tissue Miniprep System (Promega). Enrichment of mRNA was performed via oligo dT-based mRNA isolation by using NEBNext® Poly(A) mRNA Magnetic Isolation Module (NEB); libraries were prepared from 1 µg of isolated total RNA. Libraries for RNASeq were prepared using the NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (NEB) in combination with NEBNext® Multiplex Oligos for Illumina® (Dual Index Primers Set1) (NEB) according to the manufacturer's instructions. Purification steps were performed using Beckman Coulter™ AMPure XP beads. Library quality was assessed using the 4200 TapeStation system (Agilent).