GSM9553648: RNA-Seq, BJ-hTert cells, KRASG12V expression, 0h induction, Rep 2; Homo sapiens; RNA-Seq
All RNA extraction was performed using the Direct-zol RNA Miniprep Kit (Zymo Research) according to the manufacturer's instructions. Briefly, cell or nuclei pellets were extracted in TRI Reagent® (Zymo Research) and stored at -80°C until required. RNA extraction was performed by adding equal volume of ethanol to TRI Reagent® before binding to bind directly to the Zymo-Spin™ Column. The column was washed according to the manufacturer's instructions and incubated with DNase I to remove any DNA contamination. Purified RNA was eluted from the column using DNase/RNase-Free Water. RNA quality and concentration was determined using 4200 TapeStation (Agilent). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification