title: ALSP6_WM
design: Post-mortem frozen brain tissue was homogenized in a Dounce homogenizer in Lysis Buffer (10_mM Tris-HCl, pH 7.4, 10_mM NaCl, 3_mM MgCl2, and 0.025% NP-40), and incubated on ice for 15_min. The suspension was filtered through a 30-_m filter to remove debris and pelleted at 500g for 5_min at 4_oC. Nuclei were washed and filtered twice with Nuclei Wash (1%_BSA in PBS with 0.2_U _l1 RNasin (Promega)). Nuclei pellets were resuspended in 500__l nuclei wash and 900 l 1.8_M sucrose. This 1,400 _l mixture was carefully layered on top of 500__l 1.8_M sucrose and centrifuged at 13,000g for 45_min at 4_oC to separate the nuclei from myelin debris. The nuclei pellet was resuspended in nuclei wash at 1,000 nuclei _l1 and filtered through a 40-_m FlowMi Cell Strainer.
identifiers.namespace: SUB15241711
identifiers.id: ALSP6_WM