GSM5652033: ORB8266-DMSO; Homo sapiens; RNA-Seq
Approximately 300,000 cells were harvested for each sample for RNA extraction. Cells were centrifuged for 5 minutes at 1500rpm, the supernatant was removed, and the cell pellets were resuspended in 300ul of Trizol. 6-hour samples were stored at -80C overnight and 24 hour samples were kept on ice, prior to RNA extraction. RNA was extracted using Direct-zol RNA Miniprep kits (Zymo Research, USA), following the manufacturer's instructions. RNA was quantified using a NanoDropTM 2000 spectrophotometer and standardised to 100ng of RNA in 50ul of nuclease free water. Library prep was performed using Nebnext Ultra II directional RNA library prep kit for Illumina with Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, USA), following the manufacturer's instructions