GSM8030989: Replicate 11, mouse mixed pool, snATAC; Mus musculus; ATAC-seq
Twenty sex-matched B6 mice were deeply anesthetized with Isoflurane, decapitated and the spinal cord was dissected out. Then, the lumbar spinal cord from L3-5 was mounted on an agarose mold and the dorsal halve was micro dissected on a vibratome (Leica Microsystems). Next, the tissue was Dounce homogenized with a loose pestle in 2ml of Lysis Buffer containing in mM: 10 Tris-Cl, 10 NaCl, 3 MgCl2; ~25 strokes, and filtered through a 70mm cell strainer, followed by a centrifugation step at 400g for 5min at 4ºC. After that, the pellet was resuspended in 2ml of Nuclei Wash and Resuspension Buffer (1X PBS, 1% BSA and RNase inhibitor 0.2U/mL) and filtered-through a 40mm cell strainer to optimize myelin and debris removal. Finally, nuclei viability was assessed by trypan blue staining Library construction and sequencing were done at the 10X Genomics Core at the University of Pittsburgh according to standard 10x workflow: https://www.10xgenomics.com/support/single-cell-atac/documentation/steps/library-prep