GSM8381347: chx_6h; Mus musculus; RNA-Seq
Cells were washed with 1xPBS and dettached with 0.05% trypsin in EDTA for 2 minutes at 37C, followed by quenching with complete growth media. Proper viability of cells prior to freezing was quantified with propidium iodide/acridine orange staining in a LUNA-FL Dual Fluorescence Cell Counter. Cells were centrifuged at 200rcf for 3 minutes. The supernatant was discaded and the pellet was resuspended in 1mL freezing media (80% complete media, 10% heat-innactivated FBS, 10% DMSO), and frozen at -80C. Cells were shipped to comercial company in dry ice for single-cell RNA barcoding (10x Genomics' Chromium) and sequencing (NextSeq 2000 instrument, Single Cell 3' v3 chemistry), according standard procedures.