GSM7439572
Total RNA was extracted with TRIzol reagent (Fisher BioReagents, BP2805-25), followed by concentration and purification with Zymo kit, including DNase treatment to remove genomic DNA. RNA integrity was assessed with RNA 6000 Nano kit (Agilent, 5067-1511) in the 2100 Bioanalyzer. rRNA was depleted with NEB Next rRNA Depletion Kit (human, mouse, rat) (E #E6310X). For RNA-seq (mRNA) libraries generation, 200 ng of total RNA was treated with NEBNext rRNA Depletion kit (New England Biolabs, E6310X) and fragmented for 10 minutes. NEBNext Ultra II Directional (stranded) RNA Library Prep (New England Biolabs, E7760L) and NEBNext Multiplex Oligos (New England Biolabs, E6442L) kits were used for cDNA synthesis and amplification for 11 PCR cycles. The concentration of cDNA libraries was determined on Qubit Fluorometer (Thermo Fisher).