GSM5067935: HMC3 replicate 3; Homo sapiens; ssRNA-seq
Cells cultured in 6-well plates were treated with 600 µl TRIzol (ThermoFisher, 15596018) directly on plate. The solution was used for total RNA extraction using Direct-zol RNA MiniPrep Kit (Zymo Research, R2050). RNA amount was quantified using Qubit RNA HS Assay Kit (ThermoFisher, Q32852) and RNA integrity was checked by the presence of 18S and 28S bands on a 2% E-Gel EX Agarose Gel (ThermoFisher, G402002). 1 µg RNA was used for library preparation using NEBNext Ultra II Directional RNA Library Prep Kit (NEB, E7760S) following manufacturer's instruction on combined usage with NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB, E7490). Library QC and sequencing was performed by the Biopolymers Facility at Harvard Medical School. Stranded poly(A) RNA-seq