GSM8816506: M.Masseter in vivo skeletal muscle tissue; Bos taurus; RNA-Seq
Skeletal muscle tissue was minced and enzymatically digested with a 0.2 % Collagenase II and 1.5 % BSA solution for 1 hour at 37 °C in a shaking waterbath. The cell solution was filtered using a 100 µm cell strainer, followed by red blood cell depletion using ACK lysis buffer. After washing the cells with PBS, the cell solution was filtered using a 40 µm cell strainer followed by a centrigugation step(650 x g for 8 min). The cell pellet was then resuspended in FACS buffer (PBS supplemented with 0.5 % BSA) and sorted for live cells using a cell viability dye (Calcein Violet 50 AM). After counting the cells with a hemocytometer, 20000 cells were used for 10X library prepartation. Library preparation was performed following the manufacturer's instructions. Library preparation was performed using the Chromium Next GEM Single cell 3' v3.1, Dual Index kit.