GSM7698428: E1; Homo sapiens; RNA-Seq
H520 cells at 48 hours after Gapmer antisense oligonucleotide transfection and CRISPR edited and non-edited clones were washed with PBS Dulbecco's Phosphate Buffered Saline (Euroclone S.p.A., Pero, Italy) and incubated with Trypsin/EDTA 1X in Phosphate Buffered Saline (Euroclone S.p.A., Pero, Italy) at 37°C for 5 min. Cells were collected and pelleted. Total RNA was isolated from cellular pellets using QIAzol Lysis Reagent and miRNeasy Mini Kit (QIAGEN, Hilden, Germany) with DNase I digestion (QIAGEN, Hilden, Germany) according to the manufacturer's instructions. RNA integrity was checked through agarose gel electrophoresis. RNA yield and A260/A280 ratio were monitored with Nabi - UV/VIS Nano Spectrophotometer MicroDigital (TWIN HELIX SRL, Rho, Italy). For gene-expression profiling studies, RNA integrity numbers were also measured using 4200 TapeStation System (Agilent Technologies, Santa Clara, CA, USA). Total RNA was purified through oligo-dT-based RNA capturing, randomly fragmented and transformed into cDNA with NEB library preparation protocol. Library preparation was performed by Novogene, UK.