title: GSM5981429: lumbar spinal cord-0 dpi [Mac_L snRNA-seq]; Macaca mulatta; RNA-Seq
library_construction_protocol: For nuclei extract, the spinal cord was dissociated and homogenized using a glass dounce tissue grinder (D8938; Sigma-Aldrich) in cold Nuclei EZ lysis buffer (EZ PREP NUC-101; Sigma-Aldrich) on ice. The lysate was then diluted with cold sucrose buffer and centrifuged for cell debris removal. Nuclei deposited on the tube wall were collected were suspended in Nuclei Suspension Buffer (NSB; consisting of 1xPBS, 0.01% BSA, and 0.1% RNase inhibitor (Clontech, Cat #2313A)). A final nuclei suspension was filtered through a 40μm cell strainer (Corning, Cat #352235) and used for libraries preparation. Nuclei were processed using Chromium Single Cell V3 Chemistry Library Kits (10xGenomics) according to the manufacturer's instructions. In brief, nuclei were partitioned into Gel Beads-in-emulsion (GEMs) on the Chromium Controller, followed by full-length cDNA generated, barcoded, and sequenced on an Illumina HiSeq 4000.