GSM9068178: SOD1-Tg model spinal cord control; Mus musculus; RNA-Seq
mice were perfused with cold HBSS and CNS was extracted and flash frozen. A small portion of CNS regions of interest was excised from the CNS tissue and enriched for myeloid cells with modified Frankenstein's protocol. Briefly, CNS tissue was homogenized in Nuclei EZ lysis buffer (Millipore Sigma) and upon incubation was filtere through 70µm filter. The suspension was centrifuged at 600g for 5 min at 4°C. The pellet was washed with nuclei wash and resuspension buffer (1x PBS, 1.0% BSA, 0.2 U/μl RNase Inhibitor) and centrifuged at 600g for5min at 4°C.Following this, Nuclei were stained for DAPI, NeuN and Olig 2 markers and sorted by Flow cytometry. Library was performed according to the manufacter's instructions (single cell 3' v3 protocol, 10x Genomics).