title: GSM7113030: Degenerative area of injured spinal cord, 30 days, rep1; Macaca mulatta; RNA-Seq
library_construction_protocol: For cell preparation, tissues were digested using a papain dissociation system according to the manufacturer's protocol (LS003126, Worthington). Spinal cords were roughly cut into small pieces and then incubated in papain solution containing DNase I at 37℃ for 1-1.5 h with occasional pipetting. Albumin-ovomucoid inhibitor solution was used to terminate the enzymatic reaction. Subsequently, the cell suspension was further pipetted to generate a single-cell suspension, which was centrifuged at 400 g for 5 min to obtain a cell pellet. Then the cell pellet was resuspended with 20% Percoll (P1644, Sigma) and centrifuged at 400 g for 10 min to remove myelin debris. The cells were resuspended in PBS with 0.04% BSA.  For nuclei extract, the spinal cord was dissociated and homogenized using a glass dounce tissue grinder (D8938, Sigma) in cold Nuclei EZ lysis buffer (NUC101-1KT, Sigma) on ice. The lysate was then diluted with cold sucrose buffer and centrifuged for cell debris removal. Nuclei deposited on the tube wall were suspended in Nuclei Suspension Buffer consisting of 1xPBS and 0.01% BSA and 0.1% RNase inhibitor (2313A, TaKaRa). Cells and 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 NovaSeq 6000.