Single-cell and single-nucleus RNAseq of the human and mouse skeletal muscle across age
Single-cell and single-nucleus RNAseq of the human and mouse skeletal muscle across age
Three 19 months old and five 3 months old male mice of C57BL/6JRj strain were obtained from Janvier labs, France.They were used to dissect hindlimb muscles for the single-cell isolation according to the following protocol https://www.protocols.io/view/single-cell-isolation-from-human-skeletal-muscle-261geyydv479/v1. Briefly, muscle tissue was minced, digested in the solution of Collagenase II (Worthington-Biochem, LS004176) and Dispase (Gibco, 17105041) and, following that, the lysate was centrifuged in the gradient of Percoll to recover a fraction with single-cells. Cell or nuclei suspension were  loaded on the Chromium Controller (10X Genomics) with a targeted cell or nuclei recovery of 8000.  10X Genomics Single Cell 3â€™ v2 or v3 Reagent Kit was used to create GEM, perform cDNA synthesis and generate sequencing libraries according to standard instructions.