GSM9864416: 2D_D0_rep3; Homo sapiens; RNA-Seq
For sample collection at selected time points, culture medium was removed, and cells were washed with PBS. Then cells were incubated with Accutase® solution (Sigma-Aldrich®, #A6964) in the case of hiPSCs or 0.25% (v/v) trypsin-EDTA (Gibco™, #R001100) in PBS for differentiated cells, both at 37ºC for 7 min. Culture medium was added to inactivate enzymatic digestion and cells were homogenized and centrifuged at 200 xg for 3 min. The cell pellet was washed with PBS and centrifuged again at 200 xg for 3 min. After removing the PBS, cell pellet was stored at -80ºC until further analysis. Total RNA was extracted using the High Pure RNA Isolation Kit (Roche, #11828665001) following the provided instructions. Libraries were prepared using Lexogen QuantSeq 3′mRNA-Seq Library Prep. Kit FWD for Illumina using standard protocols. Sequencing was performed using Illumina HiSeq (50 cycle's protocol) or NextSeq (75 cycle's protocol) platforms