Illumina HiSeq 4000 paired end sequencing; Transposable element expression at unique loci in single cells with CELLO-seq
Transposable element expression at unique loci in single cells with CELLO-seq
For harvesting and generating single cell suspensions, human iPSCs were harvested using Accutase (Millipore, product code SCR005). Just prior to FACS sorting, cells were resuspended in 1x PBS and strained (Cell strainer (40uM), Corning #CLS431750-50EA). Single cells were sorted into 96-well plates (Framestar #4TI-0960) containing 2 µl lysis buffer with 1U/µl Rnase Inhibitor (RRI), ERCC spike in mix at 1 in 20 Mio dilution (Ambion Cat #4456740). Plates were sealed (Thermo Fisher Scientific #AB0626) immediately spun down and frozen at −80 °C until further processing.  no growth protocol  Human-induced pluripotent stem cells were thawed and cultured under feeder-free conditions on vibronectin coated 15 cm tissue culture treated plates (Corning) in complete medium (500ml Essential 8 medium ((Life Technologies #A1517001), 10 ml Essential 8 supplement, 5ml Penicillin/Streptomycin (Invitrogen #15140122)). Cells were media changed every day except for the day of passaging. Cells were incubated at 37°C, 5% CO2 and routinely passaged using 0.5 mM EDTA (LifeTechnologies #AM9260G).  no treatment We prepared cDNA of human iPSCs following the Smart-seq2 protocol.
Experimental Factor: cell type
induced pluripotent stem cell
Experimental Factor: single cell identifier
human_iPSCs_cell_shortread_66