GSM8830141: Skeletal muscle, satellite cell depleted 2, 28 month old; Mus musculus; RNA-Seq
Single cell suspensions were made from fresh muscle using collagenase followed by collagenase and dispase. Suspended cells were then filtered through a 40μm strainer and pelletted by centrifugation at 500 x g for 5 minutes and resuspended in dPBS with 0.04% BSA to reduce ambient RNA and cell aggregation per 10X Genomics recommendation. Debris, doublets, and dead cells were gated out during fluorescence activated cell sorting (FACS) using forward/side-scatter profiles and DAPI fluorescent intensity. Cells were sorted directly into reverse transcription buffer. After loading into the 10X Chromium Controller, the Single Cell 3' reagent kit was applied according to the manufacturer's protocol. Libraries were prepared using version 3.0 chemistry.