GSM7733754: Brain, control biol rep 1; Mus musculus; RNA-Seq
Flash frozen tissues were received at Azenta, South Plainfield, NJ, USA in dry-ice and stored upon receipt -80°C until further processing. Nuclei extraction was performed using the Miltenyi Nuclei Extraction Buffer (Miltenyi Biotec, Auburn, CA, USA) following manufacturer's guidelines with gentle MACS Dissociation and C tubes. Upon isolation, the nuclei were counted using trypan blue with Thermo Fisher Countess III automated cell counter 3' RNA Library preparation and Sequencing Single nuclei RNA libraries were generated using the Chromium Single Cell 3' kit (10X Genomics, CA, USA). Loading onto the Chromium Controller was performed to target capture of ~3,000 GEMs per sample for downstream analysis and processed through the Chromium Controller following the standard manufacturer's specifications. The sequencing libraries were evaluated for quality on the Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified using Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA). Pooled libraries were quantified using qPCR (Applied Biosystems, Carlsbad, CA, USA) prior to sequencing using Illumina 2 x 150bp at a target death of 50,000 pair-end end reads per cell at a configuration compatible with the recommended guidelines as outlined by 10X Genomics. snRNA-seq