GSM8478521: 0069_Healthy-A2E_5umol_L_3d_QD-No light; Homo sapiens; RNA-Seq
Total RNA was extracted using phenol chloroform-based extraction and subsequently isolated with the MagMAX-96 Total RNA Isolation Kit (Thermo Fisher Scientific, AM1830), with DNase digestion prior to final elution (Qiagen, 79254). The library was constructed with 250ng of total RNA using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina #E7760, the NEBNext Poly(A) mRNA Magnetic Isolation Module #E7490 upstream and the NEBNext Multiplex Oligos for Illumina #E7600 downstream (all New England Biolabs). The only deviation from the manufacturer's protocol was the use of Ampure XP beads (Beckman Coulter) for double-stranded cDNA purification, instead of SPRIselect Beads.