GSM7623437: hiPSC 190 PBMC 4F SeV, day 7 NOTO transfected; Homo sapiens; RNA-Seq
Cell were gently dissociated using TryplE for 7min at 37°C and then labelled using 10X CellPlex protocol for tag assignment (CG000388, 10X Genomics) 40,000 freshly collected cells were loaded into a Chromium Controller (10X Genomics), containing 5,000 untransfected cells, 10,000 cells NOTO transfected, 5,000 cells NOTO transfected and supplemented with 5µM SB431542 from day 2, repeated for two independant experiments with hiPSC lines a and b. In brief, single cells were encapsulated into Gel Beads-in-emulsion (GEM) by 10X Chromium Single Cell Controller. Single Cell 3' Reagent Kit was used to perform reverse transcription, cDNA cleanup and amplification. The double-stranded cDNA underwent enzymatic fragmentation, adapter ligation, index PCR and SPRIselect size selection as per manufacturer's protocol. All libraries were sequenced using paired-end sequencing using the Illumina NovaSeq 6000 following manufacturer's instructions.