GSM6892046: QSC; Mus musculus; RNA-Seq
QSCs and ASCs from the TA muscles of Pax7-CreERT2;Rosa26-mTmG mice were fluorescently sorted as previously described (Li et al., 2019). Briefly, mononuclear muscle-derived cells were isolated from the hind-limb muscles of adult (2-month-old) mice by digesting the muscle chops with 2 U/ml dispase and 0.2% collagenase for 45 min. The cells were filtered through 70-µm and 40-µm cell strainers, suspended at 103 –107 cells/ml, and directly sorted with a BD Aria II Cell Sorting System. QSCs and ASCs were washed and resuspended in PBS supplemented with 0.04% w/v BSA. Library was performed according to the manufacter's instructions (single cell 3' v2 protocol, 10x Genomics). Cell suspensions were loaded on a Chromium Controller instrument (10x Genomics) to generate single-cell gel bead-in emulsions (GEMs). GEM-reverse transcription (GEM-RT) was performed in a Veriti 96-well thermal cycler. After RT, GEMs were harvested and the cDNAs were amplified and cleaned with an SPRIselect Reagent Kit (Beckman Coulter). Indexed sequencing libraries were constructed using a Chromium Single-Cell 3' Library Kit (10x Genomics) for enzymatic fragmentation, end-repair, A-tailing, adaptor ligation, ligation cleanup, sample index PCR, and PCR cleanup. The libraries were sequenced with an Illumina NovaSeq 6000.