GSM7697560: Control, 4 pooled samples; Mus musculus; RNA-Seq
Cells isolated from left ventricular tissue using the gentleMACS Octo-Dissociator with heaters, gentleMACS C-tubes and Multi Tissue Dissociation Kit 2 (Miltenyi Biotec, #130-110-203). After digestion, the enzymatic reaction was quenched with DMEM (Dulbecco's medium, 20% FBS, 1% penicillin/streptomycin) and the cell suspension was passed through a 70 µm cell strainer to remove cardiomyocytes and small fragments. Cells were spun at 600g for 5 min at room temperature. Debris was then removed using Miltenyi Biotec Cell Debris Removal Solution (Cat #: 130-109-398). After debris removal, red blood cells were lysed in ACK lysis buffer (Invitrogen) for 5 min at room temperature and resuspended in PBS containing 1% BSA. Chromium Next GEM Single Cell 3' HT Kit v3.1 (10x Genomics, #1000370) according to the manufacturer's protocol. Libraries were checked for quality on an Agilent 4200 TapeStation (D1000 ScreenTape) and quantified using QuantStudio 12k Flex (Collibri library quantification kit).