GSM7157740: snRNA_seq_Sham_2; Mus musculus; RNA-Seq
In this study, for each library, the two right cortices from two mice in the same group were dissected and pooled together for the further nuclei suspension. preparation. After washed by cold 1x PBS, the brain tissue was transferred into the dounce homogenizer containing 5 ml of lysis buffer. After 20 times of up-and-down manual strokes on ice, the homogenized sample was transferred into a new 15 ml conical tube, followed by the centrifugation (4000 r.p.m., 4℃，5min). After centrifugation, the supernatant was discarded, and the precipitation (nuclei pellet) was retained. Then, the 15 ml tube was added with 1 ml of lysis buffer, and the nuclei pellet was pipetted by using a sterile 1 ml pipette tip until the nuclei were well distributed. After the nuclei suspension was added with 4 ml of lysis buffer and 5 ml of sucrose solution sequentially, the nuclei suspension was mixed well by gently inverting the tube ten times. After the centrifugation again (4000 r.p.m., 4℃，5min), the supernatant was discarded, and the precipitation (nuclei pellet) was retained. After the nuclei pellet was resuspended in 1 ml of FBS buffer, the nuclei pellet was pipetted by 1 ml and 200 μl pipette tip until the nuclei were well distributed. After 500 μl of nuclei suspension was taken for being filtered through a 40 μm cell strainer, the nuclei suspension were added 4 ml of FBS buffer. After the first nuclei count, all the nuclei solution was adjusted to a final concentration of 500-1200/μl. The single-nuclei RNA-seq libraries were prepared using Chromium Single Cell 3ʹ Reagent Kits v3 from 10x Genomics  Single nuclei RNA-seq was performed on DNBSEQ-T7.