GSM9616439: Human_CellLines_UFM1_CYTO_DMSO_ANS_rep3; Homo sapiens; RNA-Seq
Cytosolic and nuclear fractions from Arabidopsis and human RKO cells were homogenized and stored in TRIzol at −80 °C until further processing. RNA was extracted using the Direct-zol RNA Microprep Kit (Zymo Research) according to the manufacturer's instructions. Neuronal cell samples stored in 300 µL of DNA/RNA Shield were also kept at −80 °C until use. RNA from total cells of these samples was extracted using the Quick-RNA Miniprep Plus Kit (Zymo Research), following the recommended protocol. DNase I (NEB) treatment was performed during RNA isolation. Polyadenylated mRNA was enriched from total RNA using oligo(dT) magnetic beads (NEBNext Poly(A) mRNA Magnetic Isolation Module), followed by strand-specific library preparation using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina. All steps, including mRNA capture, first- and second-strand synthesis, end repair, adaptor ligation, and library amplification, were performed according to the manufacturer's instructions. Library quality and fragment size distribution were assessed using an Agilent Fragment Analyzer.