GSM8752700: MG6-4_day4_2_INFg_1; Homo sapiens; RNA-Seq
Cultured cells were dissolved directly with TRIzol (Thermo Fisher Scientific, 15596018) for total RNA purification with Direct-zol RNA MiniPrep Kit (Zymo Research, R2050). RNA concentration was quantified with Qubit RNA HS Assay Kit (Thermo Fisher Scientific, Q32852). RNA integrity was confirmed by presence of 18S and 28S bands on a 2% E-Gel EX Agarose Gel (Thermo Fisher Scientific, G402002). Between 100 ng to 1000 ng total RNA was used as input for mRNA enrichment using NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, E7490), followed by library construction with NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs, E7760S) following the manufacturer's instructions. Biopolymers Facility at Harvard Medical School performed library QC and sequencing.