GSM5625333: Day 9-RA+BMP4; Mus musculus; RNA-Seq
After a quick PBS wash at day 9 of the differentiation, cells were dissociated with 0.25% cold trypsin. Cells were pelleted and dead cells were removed using Dead Cell Removal Kit (MIltenyi Biotec). Live cells were counted using hemocytometer and resuspended in 0.04% BSA/PBS solution at 1000 cells/ ul density and library preparation and sequencing was perfomed following 10X genomics chromium protocols. Single cell suspension from day 9 cultures were prepared using 10x Genomics Chromium Single Cell 3' Reagent Kits v3 (10X Genomics, Pleasanton, CA) according to manufacture's protocol. Quantity and quality of cDNA were assessed by Agilent 2100 expert High Sensitivity DNA Assay. cDNA samples were sequenced on 1 lane of NovaSeq 6000 S2 flowcell at UCLA Technology Center for Genomics and Bioinformatics.