title: RNA-seq of Sprague Dawley rat: female adrenal gland
design: A total amount of 3 g RNA per sample was used for RNA-seq library construction. Sequencing libraries were generated using NEBNext UltraTM Directional RNA Library Prep Kit from Illumina (NEB, USA) following manufacturers protocol. Briefly, mRNA was purified and enriched from total RNA using poly-T oligo-attached magnetic beads. Purified mRNA was fragmented using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5). First-strand cDNA was generated using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H). Second strand cDNA was synthesized subsequently using DNA polymerase I and RNase H. The library fragments were purified with AMPure XP system (Beckman Coulter, Beverly, USA) to select cDNA fragments with approximately 200 bp in length. Then size-selected fragments were PCR amplified. Finally, the library was sequenced in a paired-end reads modus of 150 bp per read using Illumina HiSeq 2000 platform.
identifiers.namespace: SUB6546585
identifiers.id: Adrenal_HS5