GSM8254517: Skeletal Muscle, Non-ischemic; Homo sapiens; RNA-Seq
Skeletal muscle tissue was collected during surgery and prepared as described in Skelet Muscle. 2020;10:19. doi: 10.1186/s13395-020-00236-3 The library was performed according to manufacturer's instructions (single cell 3' v3 protocol, 10x Genomics). Briefly, cells were checked for viability before the cells were resuspended in the master mix and loaded with partitioning oil and gel beads to generate the gel-bead-in-emulsion (GEM). Poly-A RNA from cell lysates in each GEM were retrotranscipted to cDNA containg a unique molecular identifier (UMI), 10x barcode and Ilumina R1 primer sequences. cDNA was pooled and cleaned with Dynabeads, amplified by PCR and fragments of the appropriate size were selected by addition of SPRIreagent. Ilumina R2 primer sequence, paired-end constructs with P5 and P7 sequnces and a sample index were additionally added during library construction.