GSM6939128: G3KO_BMP4_6h_scRNA; Homo sapiens; RNA-Seq
Cells were washed with DPBS and dissociated with Gentle Cell Dissociation Buffer (Stem Cell Technologies) for a maximum of 20 minutes at 37 degrees, then triturated in mTeSR1. Cells were pelleted and resupended in 1% BSA in DPBS, and filtered using a 35 um cell strainer. The quality and concentration of each single cell suspension was measured using Trypan blue and the Eve automatic cell counter. Where necessary samples were diluted to a concentration of ~1000 cells/ul and approximately 10 000 cells were loaded for each sample into a separate channel of a Chromium Chip B for use in the 10X Chromium Controller. The cells were partitioned into nanoliter scale Gel Beads in emulsions (GEMs) and lysed using the Chromium Single cell 3' v3 GEM, Library and Gel Bead Kit (cat: 1000075). The RNA was reversed transcribed and amplified using 11 cycles of PCR. Libraries were prepared from the cDNA using a further 12 cycles of amplification and QC'd using the Agilent TapeStation. The libraries were pooled and sequenced on the HiSeq 4000 10x Chromium Single Cell 3' Capture v3