GSM7660054: GE_p38i_rpl1__FBC; Homo sapiens; RNA-Seq
24h after the editing treatment, viable CD34+ cells (negativite to ZombieAqua viable dye) were sorted according to surface expression of CD34+CD133+CD45RA-CD90+GFP+ and CD34+CD133+CD45RA-CD90+GFP-. Sorted population where stained for 30' at 4°C with Total Seq-B hashtag antibodies (H3 for CD34+CD133+CD45RA-CD90+GFP+ GE DMSO and GE p38i; H4 for CD34+CD133+CD45RA-CD90+GFP- GE DMSO and H5 for CD34+CD133+CD45RA-CD90+GFP- GEp38i) and viable cells were counted with Trypan Blue solution 0.4% (GIBCO) and 5x104 viable cells (2,5x104 from each population) were utilized for the subsequent procedure (estimated recovery: 3x104 cells/sample). Briefly, single cells were partitioned in Gel Beads in Emulsion (GEMs) and lysed, followed by RNA barcoding, reverse transcription and PCR amplification (11 cycles). scRNA-Seq libraries were prepared according to the manufacturer's instructions, checked and quantified on LabChip GX Touch HT (Perkin Elmer) and Qubit 3.0 (Invitrogen) instruments.