GSM8187474: G3; Mus musculus; RNA-Seq
The mouse hearts were flushed with saline, minced up and enzyme digested (2.5mg/ml collagenase I).Next, cells were passed over a 40µm cell strainer, cell debris was removed using a cell debris removal kit and cells were resuspended in red blood cell lysis buffer.  The, cells were filtered through a 30µm and 20µm filter to remove cardiomyocytes. Next, CD45+ cells were depleted using anti-CD45 anti-mouse microbeads (Miltenyi, 130-052-301) and magnetic associated cell sorting (MACS). Finally, cells were resuspended in PBS + 0.04% filtered BSA and counted using an automated cell counter (Luna FL). Samples were further diluted to achieve approx. 1x106 cells/ml (~1000 cells/µl) before loading into 10x Chromium. Cells were loaded into 10X Chromium followed by library preparation targeting an average of 10 000 cells per library. All libraries were dual index and prepared using 10X 3'kitv3.1.