GSM9682372: dFLEx CDS tumors_untreated; Mus musculus; RNA-Seq
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 AO/PI dye on the Nexcelom Cellaca MX. Single nuclei RNA libraries were generated using the Chromium Single Cell 3' kit using on chip multiplexing (10X Genomics, CA, USA). Loading was performed to target capture of ~5,000 GEMs per sample for downstream analysis, and samples were 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 loading onto an Illumina sequencing platform. The sequencing libraries were multiplexed and clustered onto a flowcell on the Illumina NovaSeq X Plus instrument according to manufacturer's instructions. The samples were sequenced at a configuration compatible with the recommended guidelines as outlined by 10X Genomics.