GSM7101940: mdx control 3; Mus musculus; RNA-Seq
Freshly dissected gastroc muscle was homogenized with homogenization buffer (10mM HEPES, 60mM KCl, 2mM EDTA, 0.5mM EGTA, 300mM sucrose, qH2O). The mixture was centrifuged for 10 minutes, resuspended in homogenization buffer, placed onto a discontinuous sucrose gradient (2.8M and 2.1M sucrose: 50mM HEPES, 25mM KCl, 5mM MgCl2, qH2O), and centrifuged for 3 hours at 4ºC. The interface was resuspended in resuspension buffer (20mM HEPES, 10mM KCl, 1.5mM MgCl2, 0.2mM EDTA, qH2O), centrifuged, and nuclei counted with BioRad TC20 Automated Cell Counter. Library was performed according to the manufacturer's instructions (single cell 3' v2 protocol, 10x Genomics)