GSM6703450: Embryonic stem cell (ES cell); Homo sapiens; RNA-Seq
Primed hESCs maintained without feeder cells were collected without any selection. Naive hESCs were sorted EOS-GFP+ (the purity was more than 95%) by using flow cytometry. AICAR-treated hESCs on MEF feeder cells were collected after MEF feeder cells (mouse-MHC+) depletion using flow cytometry. Each cell was determined by FlowCount beads (Beckman Coulter). For scRNA-seq, cDNA libraries and hashtag libraries were prepared combined by an BD Rhapsody system and an optimized process similar to the magnetic bead-based bulk RNA-seq method (Shichino et al. JCI insight 2019). Sequencing libraries were generated by using NEBNext UltraII FS library prep kit (New England Biolabs), and QC of cDNA/Hashtag libraries was performed by MultiNA (Shimazu) and qPCR library quantification assay (KAPA). BD Rhapsody WTA