GSM5322214: HepG2 S3 rep3 [I3]; Homo sapiens; RNA-Seq
Total RNAs were extracted from cells inactivated with 1 ml of TRIzol reagent. For each phase separation, 0.2 ml of chloroform was added and the aqueous phase was collected in fresh tubes and mixed with 0.5 ml of isopropyl alcohol for pelleting RNA. RNA pellets were washed with 75% ethanol and digested with DNase for 30 min to remove DNA contamination, then pelleted via addition of NaOAC and LiCl. RNA pellets were then washed with 75% ethanol and reconstituted in nuclease-free water. RNAs were reverse transcribed into cDNAs and PromethION library preparation was performed based on the manufacturer's instructions for the barcoding cNDA (SQK-LSK109, EXP-NBD104 and EXP-NBD114).