title: GSM5420220: sham2; Mus musculus; OTHER
library_construction_protocol: Nuclei were isolated from the nRT/thalamus. Tissue samples were homogenized with 10 strokes of the loose "A" pestle and 15 strokes of the tight "B" sized pstle. The lysate was passed through a 40uM FlowMi strainer and nuclei were peleted at 500 RCF at 4C. Pelleted nuclei were resuspended in the homogenization buffer, purified using an iodixanol gradient and immidiately used for snRNA-seq. SnRNA-seq libraries were processed using the Chromium Next GEM Single Cell 3'v3 library kit with dual indexing according ot the manufactuere's specifications. The chromium was loaded with 9,9000 nuclei.One GEM rxn was used for each sample. Replicate 1 and 2 nuclei were processed on different chromium runs.  Libraries were pooled based on their molar concentrations and sequenced on an Illumina NovaSeq 6000 system using an S1 flow cell and v1 300 cycle Reagent kit with 28 cycles for read 1, 90 cycles for read 2, 10 cycles for index1 (i7) and 10 cycles for index2 (i5).