title: GSM8394041: Control, baseline, snRNAseq; Mus musculus; RNA-Seq
library_name: GSM8394041
library_construction_protocol: 2-3 mm^3 sample, including the corpus callosum, was dissected from frozen mouse brain. Samples were homogenized on ice to extract nuclei using a glass Dounce homogenizer. Nuclei stained with DAPI and PI were purified via fluorescence activated nuclei sorting. Purified nuclei were collected to generate droplet-based, single nuclei RNA seq libaries using the 10X Genomics 3' Dual Index kit (v3.1). Libraries were generated using the 10X Genomics 3' Dual Index kit (v3.1) per manufacturer's instructions. In brief, nuclei were loaded onto a Chromium Chip G and partitioned into gel-bead emulsions using the 10X Chromium Controller. Poly-A mRNA captured from nuclei-bead emulsions were retrotranscribed into cDNA using poly(dT) primers and tagged with an Illumina R1 primer sequence, 10X barcode, and unique molecular identifier. cDNA was cleaned using Silane Dynabeads and was amplified via PCR. cDNA underwent size selection and fragmentation, followed by end repair, A-tailing, and adapter ligation to add P5 and P7 sequences, and i7 and i5 sample indeces to enable dual index, pair-end sequencing. Libraries were sequenced via 50nt paired-end sequencing on an Illumina NovaSeq 6000 using the following sequencing parameters: R1, 28 cylces; R2, 90 cycles; i7 index, 10 cycles; i2 index, 10 cycles. single nuclei RNA seq