title: Single cell RNA-seq analysis of skeletal muscle cells from Prrx1-ERT; tdTomato mice
design: Human quadriceps (Q) were removed remaining nerves, blood vessels, tendons and fat using fine-tipped forceps. Then transfer trimmed muscle into a new 60mm tissue dish and mince them thoroughly using the curved scissors. Add 4ml Ham's F10 Nutrient mix (Gibco) containing collagenase II (0.2% (W/V), Gibco), dispase II (2.4 U/ml, Sigma), and CaCl2 (2.5 mM, Sigma) per g of tissue into the dish and incubated at 37C for 60 min. Pass digested muscles through the 18 gauge needle five to seven times using a sterile syringe. Filtered digested slurry through a 70 m cell strainer (Falcon) and the remaining cells were washed and pelleted by centrifugation at 500g for 5 min. Resuspend the pellet in FACS buffer and adjust cell concentration to 1107 cells/ml. Single cell libraries was created using the 10X Genomics Chromium Single Cell 3' Library Kit.